Phaidra - University of Veterinary Medicine Vienna
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Jahresbericht 2023 / Messerli Forschungsinstitut
Der Jahresbericht gewährt Einblicke und Rückblicke in das Institutsleben und fasst Informationen zu Forschung und Lehre am Messerli Forschungsinstitut zusammen. Der Bericht liegt in deutscher und englischer Sprache vor
The myxozoans Myxobolus cerebralis and Tetracapsuloides bryosalmonae modulate rainbow trout immune responses: quantitative shotgun proteomics at the portals of entry after single and co-infections
Little is known about the proteomic changes at the portals of entry in rainbow trout after infection with the myxozoan parasites, Myxobolus cerebralis, and Tetracapsuloides bryosalmonae. Whirling disease (WD) is a severe disease of salmonids, caused by the myxosporean M. cerebralis, while, proliferative kidney disease (PKD) is caused by T. bryosalmonae, which instead belongs to the class Malacosporea. Climate change is providing more suitable conditions for myxozoan parasites lifecycle, posing a high risk to salmonid aquaculture and contributing to the decline of wild trout populations in North America and Europe. Therefore, the aim of this study was to provide the first proteomic profiles of the host in the search for evasion strategies during single and coinfection with M. cerebralis and T. bryosalmonae.One group of fish was initially infected with M. cerebralis and another group with T. bryosalmonae. After 30 days, half of the fish in each group were co-infected with the other parasite. Using a quantitative proteomic approach, we investigated proteomic changes in the caudal fins and gills of rainbow trout before and after co-infection.In the caudal fins, 16 proteins were differentially regulated post exposure to M. cerebralis, whereas 27 proteins were differentially modulated in the gills of the infected rainbow trout post exposure to T. bryosalmonae. After co-infection, 4 proteins involved in parasite recognition and the regulation of host immune responses were differentially modulated between the groups in the caudal fin. In the gills, 11 proteins involved in parasite recognition and host immunity, including 4 myxozoan proteins predicted to be virulence factors, were differentially modulated.The results of this study increase our knowledge on rainbow trout co-infections by myxozoan parasites and rainbow trout immune responses against myxozoans at the portals of entry, supporting a better understanding of these host-parasite interactions
Biological and physico-chemical properties of the soil in an acidified environment influenced by previous mining activities at the Šobov locality (SLOVAKIA)
More than 20 years after remedial measures were carried out, six plots (S1 - S6) at the Sobov locality were examined in more detail. From a pedological viewpoint, the physico-chemical differences of the soils in these areas are not the result of pedogenesis. This is the effect of extremely acidic mineralised solutions that have leaked from or are still leaking from the heap at the site. The plant community here is most often poor to weakly developed with acidophilic vegetation (S1, S2, S5), without vegetation (S4) or with dense species-rich vegetation (S3, S6). Saprotrophic microscopic fungi of the phylum Zygomycota are notably suppressed and their diversity is low. Species of the genera Absidia and Zygorhynchus were found most often. In contrast, the diversity of the phylum Ascomycota is notably high. The genera Penicillium (35 species), Aspergillus (7 species) and Trichoderma (5 species) are dominantly represented. Species of microscopic filamentous fungi in every soil sample that did not occur in the other samples were also recorded; i.e. they form the specific soil mycobiome of the given location. From the 15 types of keratinophilic fungi, the most commonly occurring were Purpureocillium lilacinum and Keithomyces carneus. Keratinolytic properties were recorded only in the species Trichophyton ajelloi. Soil reaction is the most important ecological factor that influences the biological properties of soils
Transcriptomic profiling of different developmental stages reveals parasitic strategies of Wohlfahrtia magnifica, a myiasis-causing flesh fly
Wohlfahrtia magnifica is an obligatory parasite that causes myiasis in several warm-blooded vertebrates. Adult females deposit the first-stage larvae directly onto wounds or natural body orifices (e.g., genitalia) of the host, from where they quickly colonize the host tissue and feed on it for development. The infestation of W. magnifica can lead to health issues, welfare concerns, and substantial economic losses. To date, little is known about the molecular mechanisms of the W. magnifica-causing myiasis.In this study, we collected parasitic-stage larvae of W. magnifica from wounds of naturally infested Bactrian camels, as well as pupae and adult flies reared in vitro from the wound-collected larvae, for investigating the gene expression profiles of the different developmental stages of W. magnifica, with a particular focus on examining gene families closely related to the parasitism of the wound-collected larvae. As key proteins related to the parasite-host interaction, 2049 excretory/secretory (ES) proteins were identified in W. magnifica through the integration of multiple bioinformatics approaches. Functional analysis indicates that these ES proteins are primarily involved in cuticle development, peptidase activity, immune response, and metabolic processes. The global investigation of gene expression at different developmental stages using pairwise comparisons and weighted correlation network analysis (WGCNA) showed that the upregulated genes during second-stage larvae were related to cuticle development, peptidase activity, and RNA transcription and translation; during third-stage larvae to peptidase inhibitor activity and nutrient reservoir activity; during pupae to cell and tissue morphogenesis and cell and tissue development; and during adult flies to signal perception, many of them involved in light perception, and adult behavior, e.g., feeding, mating, and locomotion. Specifically, the expression level analysis of the likely parasitism-related genes in parasitic wound-collected larvae revealed a significant upregulation of 88 peptidase genes (including 47 serine peptidase genes), 110 cuticle protein genes, and 21 heat shock protein (hsp) genes. Interestingly, the expression of 2 antimicrobial peptide (AMP) genes, including 1 defensin and 1 diptericin, was also upregulated in the parasitic larvae.We identified ES proteins in W. magnifica and investigated their functional distribution. In addition, gene expression profiles at different developmental stages of W. magnifica were examined. Specifically, we focused on gene families closely related to parasitism of wound-collected larvae. These findings shed light on the molecular mechanisms underlying the life cycle of the myiasis-causing fly, especially during the parasitic larval stages, and provide guidance for the development of control measures against W. magnifica
Assessment of stress levels in trotting horses in conjunction with age, performance and management
Diploma thesis - University of Veterinary Medicine Vienna - 2024Young Standardbred racehorses start their careers as two-year-old, when their muscular-skeletal system is still growing. Racehorses end their careers earlier than most other performance horses. They have injuries related to the high speeds they travel and the intensity of the training as well as the repercussions from stress, such as a high prevalence of gastric ulcer. It is not completely understood whether this is caused by the early and demanding start into training, but research indicates that there is no positive correlation, and that early training might even be beneficial for a longer and sounder career. However, it remains unclear how this affects stress levels and performance of young racehorses. We hypothesised that young horses that start training early show more signs of stress compared to older horses and that stress negatively affects the performance of Standardbred racehorses. Horses (n = 45) of different age groups (two to eleven years) from three successful German Standardbred trainers were assessed by their behavior towards humans in the stall, their acceptance of harnessing and their willingness in training. This assessment was combined with a salivary cortisol concentration measurement and correlated with the horses’ performance. In this study no difference was found between the age groups regarding salivary cortisol concentrations and behavior. There was a significant difference between the three trainers regarding the behavior assessment, but this did not affect the horses’ performance. We conclude that young Standardbred racehorses appear to have similar stress levels as older ones have whereas environmental factor play a more important role than age, since horses in quieter, stables in rural areas with more pasture time showed less signs of stress.Diploma thesis - University of Veterinary Medicine Vienna - 2024Young Standardbred racehorses start their careers as two-year-old, when their muscular-skeletal system is still growing. Racehorses end their careers earlier than most other performance horses. They have injuries related to the high speeds they travel and the intensity of the training as well as the repercussions from stress, such as a high prevalence of gastric ulcer. It is not completely understood whether this is caused by the early and demanding start into training, but research indicates that there is no positive correlation, and that early training might even be beneficial for a longer and sounder career. However, it remains unclear how this affects stress levels and performance of young racehorses. We hypothesised that young horses that start training early show more signs of stress compared to older horses and that stress negatively affects the performance of Standardbred racehorses. Horses (n = 45) of different age groups (two to eleven years) from three successful German Standardbred trainers were assessed by their behavior towards humans in the stall, their acceptance of harnessing and their willingness in training. This assessment was combined with a salivary cortisol concentration measurement and correlated with the horses’ performance. In this study no difference was found between the age groups regarding salivary cortisol concentrations and behavior. There was a significant difference between the three trainers regarding the behavior assessment, but this did not affect the horses’ performance. We conclude that young Standardbred racehorses appear to have similar stress levels as older ones have whereas environmental factor play a more important role than age, since horses in quieter, stables in rural areas with more pasture time showed less signs of stress.Diplomarbeit - Veterinärmedizinische Universität Wien - 202
Comparison of a multiplex PCR with DNA barcoding for identification of container breeding mosquito species
Identification of mosquitoes greatly relies on morphological specification. Since some species cannot be distinguished reliably by morphological methods, it is important to incorporate molecular techniques into the diagnostic pipeline. DNA barcoding using Sanger sequencing is currently widely used for identification of mosquito species. However, this method does not allow detection of multiple species in one sample, which would be important when analysing mosquito eggs. Detection of container breeding Aedes is typically performed by collecting eggs using ovitraps. These traps consist of a black container filled with water and a wooden spatula inserted for oviposition support. Aedes mosquitoes of different species might lay single or multiple eggs on the spatula. In contrast to Sanger sequencing of specific polymerase chain reaction (PCR) products, multiplex PCR protocols targeting specific species of interest can be of advantage for detection of multiple species in the same sample.For this purpose, we adapted a previously published PCR protocol for simultaneous detection of four different Aedes species that are relevant for Austrian monitoring programmes, as they can be found in ovitraps: Aedes albopictus, Aedes japonicus, Aedes koreicus, and Aedes geniculatus. For evaluation of the multiplex PCR protocol, we analysed 2271 ovitrap mosquito samples from the years 2021 and 2022, which were collected within the scope of an Austrian nationwide monitoring programme. We compared the results of the multiplex PCR to the results of DNA barcoding.Of 2271 samples, the multiplex PCR could identify 1990 samples, while species determination using DNA barcoding of the mitochondrial cytochrome c oxidase subunit I gene was possible in 1722 samples. The multiplex PCR showed a mixture of different species in 47 samples, which could not be detected with DNA barcoding.In conclusion, identification of Aedes species in ovitrap samples was more successful when using the multiplex PCR protocol as opposed to the DNA barcoding protocol. Additionally, the multiplex PCR allowed us to detect multiple species in the same sample, while those species might have been missed when using DNA barcoding with Sanger sequencing alone. Therefore, we propose that the multiplex PCR protocol is highly suitable and of great advantage when analysing mosquito eggs from ovitraps
Electrical impedance tomography in anaesthetised chickens (Gallus domesticus)
The applicability of electrical impedance tomography (EIT) in birds is unknown. This study aimed to evaluate the use of EIT in anaesthetised chickens in four recumbency positions. Four adult Hyline chickens were anaesthetised with isoflurane in oxygen, and intubated endotracheally for computed tomography (CT). A rubber belt was placed around the coelom caudal to the shoulder joint. A chicken-specific finite element (FE) model, which is essential to generate anatomically accurate functional EIT images for analysis, was constructed based on the CT images obtained at the belt level. Ten additional chickens were anaesthetised with the same protocol. An EIT electrode belt was placed at the same location. The chickens were breathing spontaneously and positioned in dorsal, ventral, right and left lateral recumbency in a randomised order. For each recumbency, raw EIT data were collected over 2 min after 13 min of stabilisation. The data were reconstructed into functional EIT images. EIT variables including tidal impedance variation (TIV), centre of ventilation right to left (CoVRL) and ventral to dorsal (CoVVD), right to left (RL) ratio, impedance change (ΔZ) and eight regional impedance changes including the dorsal, central-dorsal, central-ventral and ventral regions of the right and left regions were analysed. Four breathing patterns (BrP) were observed and categorised based on the expiratory curve. A linear mixed model was used to compare EIT variables between recumbencies. Fisher\u27s exact test was used to compare the frequencies of breathing patterns for each recumbency. The ΔZ observed was synchronous to ventilation, and represented tidal volume of the cranial air sacs as confirmed by CT. Significant differences were found in CoVVD and regional impedance changes between dorsal and ventral recumbencies (P< 0.05), and in CoVRL, RL ratio and regional impedance changes between right and left recumbencies (P< 0.05), which suggested a tendency for the distribution of ventilation to shift towards non-dependent air sacs. No differences were found for TIV and respiratory rate between recumbencies. Recumbency had a significant effect on the frequencies of each of the four BrPs (P = 0.001). EIT can monitor the magnitude and distribution of ventilation of the cranial air sacs in different recumbencies in anaesthetised chickens
Cat and dog owners\u27 expectations and attitudes towards advanced veterinary care (AVC) in the UK, Austria and Denmark
Modern veterinary medicine offers a level of care to cats and dogs similar to that available to their owners, including blood transfusions, chemotherapy and MRI scans. The potential benefits to the animals of owners who can afford such care are obvious, but there can also be negative consequences if owners with strong emotional attachments to their pets pursue treatments that significantly reduce the quality of the animal\u27s life while attempting to prolong it. Moreover, caring for a chronically or seriously ill animal can lead to emotional distress and financial and practical challenges for the pet owner. A questionnaire was used to survey cat and dog owners from representative samples of citizens in the UK, Austria and Denmark, to investigate owners\u27 expectations and attitudes towards advanced veterinary care, and the factors that might influence those views. Overall, 58.4% of the pet owners surveyed believed that their pets should have access to the same treatment options as humans, while 51.5% believed that they should have access to the same diagnostic tests as humans. Owners were most likely to be neutral on the question of whether advanced veterinary care has \u27gone too far\u27 (45.3%), and to disagree with the statement that advanced care is \u27unnecessary\u27 (40.1%). In all three countries, the level of attachment owners had to their pets was most strongly associated with attitudes towards advanced care, with owners scoring higher on Lexington Attachment to Pets Scale (LAPS) being more likely to expect advanced care to be available. Other factors such as owner age, living situation (alone or not), income or possession of pet insurance were less consistently with owner attitudes. Our findings will help inform veterinarians and other health care providers about pet owner expectations and attitudes towards advanced veterinary care, and contribute to the debate on increasing specialisation within the profession
Detection of mcr-1-1 Positive Enteropathogenic Escherichia coli Isolates Associated with Post-Weaning Diarrhoea in an Organic Piglet-Producing Farm in Austria
Postweaning diarrhoea (PWD) is a frequent multifactorial disease occurring in swine stocks worldwide. Since pathogenic Escherichia (E.) coli play a pivotal role in the pathogenesis of PWD and porcine E. coli are often resistant to different antibiotics, colistin is frequently applied to treat piglets with PWD. However, the application of colistin to livestock has been associated with the emergence of colistin resistance. This case report describes the detection of the colistin resistance gene mcr-1-1 in two E. coli isolated from piglets with PWD in an Austrian organic piglet-producing farm, which was managed by two farmers working as nurses in a hospital. Both mcr-1-positive E. coli were further analysed by Illumina short-read-sequencing, including assemblies and gene prediction. Both isolates belonged to the same clonal type and were positive for eaeH and espX5, which are both virulence genes associated with enteropathogenic E. coli (EPEC). Due to the detection of mcr-1-positive EPEC and based on the results of the antimicrobial resistance testing, the veterinarian decided to apply gentamicin for treatment instead of colistin, leading to improved clinical signs. In addition, after replacing faba beans with whey, PWD was solely observed in 2/10 weaned batches in the consecutive months
Mycoplasma agalactiae Vaccines: Current Status, Hurdles, and Opportunities Due to Advances in Pathogenicity Studies
Contagious agalactia (CA) is a serious multietiological disease whose classic etiological agent is Mycoplasma agalactiae and which causes high morbidity and mortality rates in infected herds. CA is classified as a notifiable disease by the World Organization for Animal Health due to its significant worldwide economic impact on livestock, primarily involving goat and sheep farms. The emergence of atypical symptoms and strains of M. agalactiae in wildlife ungulates reestablishes its highly plastic genome and is also of great epidemiological significance. Antimicrobial therapy is the main form of control, although several factors, such as intrinsic antibiotic resistance and the selection of resistant strains, must be considered. Available vaccines are few and mostly inefficient. The virulence and pathogenicity mechanisms of M. agalactiae mainly rely on surface molecules that have direct contact with the host. Because of this, they are essential for the development of vaccines. This review highlights the currently available vaccines and their limitations and the development of new vaccine possibilities, especially considering the challenge of antigenic variation and dynamic genome in this microorganism