Phaidra - University of Veterinary Medicine Vienna
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Retrospective Screening for Zoonotic Viruses in Encephalitis Cases in Austria, 2019-2023, Reveals Infection with Lymphocytic Choriomeningitis Virus but Not with Rustrela Virus or Tahyna Virus
No full textZoonotic viruses may be neglected as etiologies of meningoencephalitis in humans. We performed retrospective testing of cerebrospinal fluid from encephalitis cases in biobank material for three zoonotic or potentially zoonotic viruses: rustrela virus (Rubivirus strelense, Matonaviridae); Tahyna virus (Orthobunyavirus tahynaense, Peribunyaviridae); and lymphocytic choriomeningitis virus ("LCMV", Mammarenavirus choriomeningitidis, Arenaviridae). The cohort consisted of 443 samples, received for routine diagnostic testing year-round between January 2019 and February 2023, and were negative for herpes simplex viruses, varicella zoster virus, and enteroviruses. Using published RT-qPCR protocols, we did not detect rustrela virus or Tahyna virus in any sample. Using a herein described RT-qPCR protocol, we detected LCMV in one sample. Partial genetic sequencing of the virus suggested that the virus was locally acquired. Our study provides information about the incidence of these viruses in humans in Austria when encephalitis is suspected
First molecular evidence of hemotropic mycoplasmas in goats from Bosnia and Herzegovina
No full textHemoplasmas represent the type of bacteria that infect red blood cells, potentially leading to various health impacts, including changes in blood parameters. The close interaction between hemoplasma and red blood cells results in cell damage through immune-related and other unspecified mechanisms. Even with a strong immune response and antibiotic treatment, affected animals are likely to remain chronic carriers once clinical symptoms have subsided. These microorganisms were previously documented in sheep and other small ruminants worldwide.Since there is a lack of research on the link between Mycoplasma infection and blood parameters, our aim was to investigate how Mycoplasma infection affects these blood parameters. In addition, the study conducted in Bosnia and Herzegovina represents the first documented research of hemoplasma infection in goats within this region.In this research, 20 Alpine goats were sampled to investigate the presence of hemoplasma using polymerase chain reaction (PCR) analysis. Sequences of the 16S rRNA gene fragments were identified subsequently. The effect of Mycoplasma ovis (M. ovis) infection was observed on the following hematological parameters: Red blood cell count (RBC), hematocrit (HCT), hemoglobin (HGB), mean cell volume (MCV), mean cell hemoglobin, mean cell hemoglobin concentration, Reticulocyte count, and white blood cell (WBC). Effect on white blood cell differentiation, absolute white blood cell counts, platelet count, and mean platelet volume were also investigated.PCR analysis confirmed the presence of Mycoplasma spp. in 7 out of the 20 blood samples. Sequencing of the 16S rRNA gene fragments revealed that all positive samples were identified as M. ovis. The research findings highlighted potential effects on blood parameters in infected goats. Goats infected with M. ovis exhibited higher mean levels of HGB and HCT compared to uninfected goats. However, there were no statistically significant differences in RBC counts between infected and uninfected groups. The study also noted significantly higher WBC counts in goats without M. ovis infection.35% of animals tested positive for M. ovis. Our study\u27s findings showed notable differences in hematological parameters between goats infected with M. ovis and those that were not infected
Anti-Cancer Potential of a new Derivative of Caffeic Acid Phenethyl Ester targeting the Centrosome
No full textAnaplastic Large Cell Lymphoma (ALCL) is an aggressive T-cell lymphoma affecting children and young adults. About 30% of patients develop therapy resistance therefore new precision medicine drugs are highly warranted. Multiple rounds of structure-activity optimization of Caffeic Acid Phenethyl Ester have resulted in CM14. CM14 causes upregulation of genes involved in oxidative stress response and downregulation of DNA replication genes leading to G2/M arrest and subsequent apoptosis induction. In accordance with this, an unbiased proteomics approach, confocal microscopy and molecular modeling showed that TUBGCP2, member of the centrosomal ?-TuRC complex, is a direct interaction partner of CM14. CM14 overcomes ALK inhibitor resistance in ALCL and is also active in T-cell Acute Lymphoblastic Leukemia and Acute Myeloid Leukemia. Interestingly, CM14 also induced cell death in docetaxel-resistant prostate cancer cells thus suggesting an unexpected role in solid cancers. Thus, we synthesized and thoroughly characterized a novel TUBGCP2 targeting drug that is active in ALCL but has also potential for other
malignancies
Preserved bevacizumab (Avastin®) eye drops for application in multidose containers - an in-vitro characterisation
No full textMonoclonal antibodies have made an immense contribution to the treatment of various human diseases. We aimed at investigating an affordable treatment option for veterinary patients with corneal neovascularization by adding the preservative benzalkonium chloride (BAC) to bevacizumab (Avastin®) for usage in multidose containers. A comprehensive analytical similarity assessment of preserved and unpreserved bevacizumab after dilution and storage was carried out.Diluted and preserved bevacizumab was analysed at different time points for a 4-week period and compared with unpreserved bevacizumab at the same concentrations at each time point. Native-PAGE, immunoblotting and HP-SEC were used to observe aggregation and degradation. DLS provided information about particle size and dispersity. Bevacizumab quantified by ELISA was conducted to determine its biological activity. Dose response curves and cell migration assays were performed to detect possible toxic effects and determine biological activity and efficacy of the drug using HUVECs.Native-PAGE, immunoblotting and HP-SEC analysis did not show any changes or degradation products in the presence of BAC and after storage compared to unpreserved bevacizumab. The overlapping intensity-based particle size distribution obtained from DLS showed similarity in all tested groups and homogeneity was maintained. ELISA accurately detected bevacizumab at different concentrations. HUVECs incubated with preserved or unpreserved bevacizumab showed a comparable effect on cell migration. No decrease in cell viability was detected.Equivalence tests demonstrated that bevacizumab is stable after dilution, storage and preservation with BAC. Our study shows that preserved bevacizumab applied in mutidose containers can be considered as a cost-effective alternative to the otherwise single-dose treatments
Metabolome and oxidative stress markers in the seminal plasma of Holstein bulls and their relationship with the characteristics of fresh and frozen/thawed sperm
No full textSeminal plasma composition has important role in sperm functionality and its freezability. The objective of this study was to test the hypothesis that seminal plasma (SP) oxidative status and metabolome are associated with fresh semen characteristics and freezability of bull sperm. To accomplish this objective, oxidative status markers and metabolome of SP of ejaculates obtained from 20 Holstein bulls (3 for each bull) were analyzed using spectrophotometry and nuclear magnetic resonance (1H NMR). The ejaculates were classified into higher motility fresh semen (HMF) and lower motility fresh semen (LMF), according to total motility (TM) and progressive motility (PM) values of fresh semen. Then the ejaculates was cryopreserved and assigned to higher motility thawed group (HMT) or lower motility thawed group (LMT) according to TM and PM at 0 h post-thawing. Multivariate analyses were performed to identify the association between the functional characteristics of fresh and thawed semen and the SP parameters, in terms of the oxidative status and the metabolomic composition. According to our results, the advanced oxidative protein products (AOPP) and thiol concentrations in SP are significantly related to some physiological characteristics of the thawed sperm, such as higher viability, TM, PM and LIN and lower mitochondrial and cytoplasmic superoxide production in viable thawed cells. In contrast, a higher amount of C in the SP was negatively related to TM and PM of thawed semen and was associated with higher mitochondrial and cytoplasmic superoxide production. In addition, partial least squares-discriminant analysis (PLS-DA) performed on the 1H NMR spectra indicated a discrete separation between HMF and LMF groups, and good discrimination between HMT and LMT groups. Higher levels of formic acid, lactate, glycerol and phosphocholine, were found in the SP of the HMF group than in the LMF group. On the other hand, alanine, phenylalanine, and tyrosine were higher in the SP of the LMF group than in the HMF group. GABA, glutamate, histidine and glycerol were found in higher concentrations in the HMT group than in the LMT group, while fructose decreased in the HMT group. Our results showed that the oxidative and metabolomic status of SP is related to the physiological properties of semen and its freezability and open new fields in research of SP biomarkers of bull semen preservation and fertility
Uncovering the full potential of attitude measures in navigating human-wolf coexistence
No full textAttitudes towards wolves are important indicators of what wolf presence means to people and whether they lean towards support or opposition. Over the past 50 years, attitude surveys and interviews have uncovered that the polarisation between social groups is not only driven by tangible impacts. Moreover, uneven distribution of intangible costs fostered feelings of marginalisation, and socio-cultural divides create different perceptions of wolves and their management. Despite these revelations, little emphasis has been placed on the intricate psychological mechanisms underlying attitude polarisation and behavioural implications. This Perspective drafts a framework around the roles of attitude strength, accessibility, and ambivalence in attitude and behaviour formation, and emphasizes the potential role of unconscious, implicit attitudes beyond explicit self-report in how humans perceive and react to wolves. It explores how these factors may explain polarisation, sudden attitude shifts, mismatches between reported attitudes and behaviours, and the unexpected ineffectiveness of some interventions. Under the assumption of this framework, approaches are discussed that could help navigate attitude shifts amid expanding wolf populations and emotional conflicts between groups and species. Though speculative for now, we hope that this bottom-up approach will guide and inspire research to further explore the proposed mechanisms and improve our understanding about how latent attitudes, ambivalence, and experiences may shape attitudes towards wolves. Rather than advocating for nationwide shifts towards positive views of wolves, we stress the importance of recognizing the full spectrum of attitudes, contexts, and social group settings in managing conflict sustainably, particularly considering expanding and increasingly urbanized wolf populations
Utilizing Noseband Sensor Technology to Evaluate Rumination Time as a Predictor of Feeding and Locomotion Behaviors in Dairy Cows
No full textDuring dental scaling in dogs under general anaesthesia, contamination of the peripheral intravenous catheter (PIVC) is unavoidable due to splatter and the generated aerosol. Bacterial contamination was compared between two commonly used PIVC placement sites. Thirty-nine client-owned dogs with a minimum length from their nose to their tail base of 50 cm were randomly assigned to receive a PIVC in either their cephalic or saphenous vein. Irrespective of the PIVC placement site, brain heart infusion agar dishes were placed in the cephalic and saphenous vein areas. Their lids were closed 0, 5, and 10 min into the procedure. Contamination was measured by counting the colony-forming units after incubation on different substrates. The data were analysed with descriptive statistics, ANOVA, and ANCOVA (p< 0.05). The cephalic vein area showed a significantly higher bacterial load than the saphenous vein area (p ≈ 0.0) regardless of the length of the dog. Furthermore, the dorsal PIVC injection ports were sampled before and after scaling, and the colonies isolated were counted and subjected to MALDI-TOF-MS for identification. The bacteria mainly belonged to the genera Staphylococcus, Neisseria, and Bacillus. Our results suggest that for dental scaling in dogs, the PIVC should be placed in the pelvic limb whenever possible to reduce the potential risk of contamination
Safe Periods and Safe Activities: Two Phenological Responses to Mortality
No full textPhenology is often thought to evolve mainly in response to food availability, yet recent studies have focused on predation. Predation may explain apparent mismatches between phenology and resources. One type of phenological response to predation involves shifting phenology from a period of high to low predation (i.e., a safe-period strategy). This strategy presupposes variation in predation over time due to environmental factors such as the number or diversity of predators. Predation varies not only over time but also among different activities like reproduction and dormancy. Alternative activities involve alternative behavioral or physiological states, and different locations where they take place influencing predation risk. Phenological responses to predation may involve shifting from a high risk activity to a safer one, resulting in increased survival (i.e., a «safe-activity» strategy). This strategy may theoretically evolve under environmental conditions associated with constant predation over time, but assumes variation in predation among activities. Safe-period and safe-activity strategies are not mutually exclusive, but assume different conditions for their evolution. On the basis of a literature review, our goal was to: (1) propose a classification of phenological responses to predation according to their evolutionary context, including mean population responses and interindividual differences (degree of synchrony); (2) to show how these two strategies may explain the lack of support for the idea that phenology responds primarily to food availability; and (3) to propose several approaches for testing the influence of predation on phenology. Our review highlights the relevance of studying phenology on multiple scales, thereby integrating several interspecific interactions (communities scales) and multiple activities (annual scale), and studying synchronicity and the pace-of-life (inter-individual scale)
Repurposing the prostaglandin analogue treprostinil and the calcium-sensing receptor modulator cinacalcet to revive cord blood as an alternate source of hematopoietic stem and progenitor cells for transplantation
No full textThe expanding field of hematopoietic cell transplantation (HCT) for non-malignant diseases, including those amenable to gene therapy or gene editing, faces challenges due to limited donor availability and the toxicity associated with cell collection methods. Umbilical cord blood (CB) represents a readily accessible source of hematopoietic stem and progenitor cells (HSPCs); however, the cell dose obtainable from a single cord blood unit is frequently insufficient. This limitation can be addressed by enhancing the potency of HSPCs, specifically their capacity to reconstitute hematopoiesis. In our study, we investigated the combined effects of treprostinil, a prostaglandin analog, and cinacalcet, a calcium-sensing receptor modulator, on the reconstitution of hematopoiesis.A Lineage Cell Depletion Kit was employed to isolate lineage-negative (lin-) HSPCs from mouse bone marrow. A Human CB CD34 Positive Selection Kit was utilized to isolate CD34+ cells from the CB of healthy donors. In vitro, the effects of treprostinil, cinacalcet, and their combination on the migration, adhesion, and differentiation of HSPCs were assessed. In vivo, homing and engraftment were examined. Eight-week-old female and male C57BL/6J, BALB/c, or female NSG mice served as recipient models.When administered concomitantly, treprostinil and cinacalcet exhibited mutual antagonism: the survival of recipient animals was lower when both drugs were administered together compared to either agent alone. Conversely, a sequential regimen involving priming with treprostinil/forskolin followed by cinacalcet treatment in vivo enhanced survival, irrespective of whether hematopoiesis was reconstituted by human or murine HSPCs. In vitro assays demonstrated enhanced migration and adhesion in response to the presence of treprostinil and cinacalcet, suggesting potential synergistic effects. Colony formation confirmed synergism.Augmenting the bone marrow reconstitution potential of HSPCs with treprostinil and cinacalcet shows promise for rescuing patients undergoing HCT. This approach is particularly beneficial for those patients at high risk of transplant failure due to limited numbers of available HSPCs. Furthermore, enhancing the potency of HSPCs has the potential to alleviate the burden and risks associated with HSPC donation, as it would reduce the number of cells needed for collection
Characterization of the colostrum proteome of primiparous Holstein cows and its association with colostrum immunoglobulin G concentrations
No full textBackground
The objective was to characterize the colostrum proteome of primiparous Holstein cows in association with immunoglobulin G (IgG) content. Immediately after calving, colostrum samples were collected from 18 cows to measure IgG concentration. Based on colostrum IgG content, samples were classified through cluster analysis and were identified as poor, average, and excellent quality. The proteome was assessed with quantitative shotgun proteomics; abundance data were compared among the colostrum types; enrichment analysis of metabolic processes and proteins classes was performed as well. We also tested correlations between this proteome and blood globulin level of cows and passive immunity level of calves.
Results
On average, 428 proteins were identified per sample, which belonged mainly to cellular process, biological regulation, response to stimulus, metabolic process, and immune system process. Most abundant proteins were complement C3 (Q2UVX4), alpha-S1-casein (P02662), Ig-like domain-containing protein (A0A3Q1M032), albumin (A0A140T897), polymeric immunoglobulin receptor (P81265), lactotransferrrin (P24627), and IGHG1*01 (X16701_4). Colostrum of excellent quality had greater (P < 0.05) abundance of serpin A3-7 (A2I7N3), complement factor I (A0A3Q1MIF4), lipocalin/cytosolic fatty-acid binding domain-containing protein (A0A3Q1MRQ2), complement C3 (E1B805), complement component 4 binding protein alpha (A0AAF6ZHP5), and complement component C6 (F1MM86). However, colostrum of excellent quality had lower (P < 0.05) abundance of HGF activator (E1BCW0), alpha-S1-casein (P02662), and xanthine dehydrogenase/oxidase (P80457). This resulted in enrichment of the biological processes predominantly for complement activation alternative pathway, complement activation, complement activation classical pathway, humoral immune response, leukocyte mediated immunity, and negative regulation of endopeptidase activity in excellent-quality colostrum. Additionally, some colostrum proteins were found to be correlated with the blood globulin level of cows and with the passive immunity level of calves (P < 0.05; r ≥ 0.57).
Conclusions
This study provides new insights into the bovine colostrum proteome, demonstrating associations between IgG levels and the abundance of other proteins, as well as the enrichment of metabolic processes related to innate immune response. Thus, results suggest that the colostrum proteomic profile is associated with the content of IgG. Future research should deeply explore the association of these findings with pre-calving nutrition status and blood composition of the cow, and with passive immunity transfer to the calf