Correlation of Faecal Egg Counts with Clinical Parameters and Agreement between Different Raters Assessing FAMACHA©, BCS and Dag Score in Austrian Dairy Sheep

Gastrointestinal nematodes, most notably trichostrongylids, are known to cause significant losses in sheep production. Previous studies have shown that monitoring parameters (e.g., FAMACHA©, BCS, dag score) change with increasing egg excretion. These parameters are well known and frequently used for targeted selective treatment. Based on the willingness to participate in this study (based on a previous questionnaire distribution among sheep farmers in Austria) we investigated the associations between faecal egg counts and the FAMACHA©, BCS, and dag scores of 1195 dairy ewes. Faecal samples were analysed using the Mini-FLOTAC technique I and larval culture. Three raters assessed the FAMACHA©, BCS, and dag scores in sheep to calculate the inter-rater agreement and intraclass correlation coefficient. The responses to the questionnaire of 23 farms were used for the evaluation, of which 16 farms were visited. Trichostrongylid eggs were detected in 95% of the faecal samples. The BCS was negatively correlated with the eggs per gram of faeces (EpG) (r = -0.156; p< 0.001) and the FAMACHA© score was slightly positively correlated with EpG (r = 0.196; p< 0.001). A small proportion of sheep (25%) shed the majority of eggs (47% to 84%). A moderate to good agreement for the parameters was found between the raters. In conclusion, the clinical parameters showed only weak correlations with faecal egg counts, and we confirmed that a minority of the flock is responsible for the majority of the pasture contamination with trichostrongylid eggs. Clinical raters should be trained before a study to increase the agreement between them

Demodicosis in a Kunekune pig and molecular characterisation of porcine demodectic mites involved: a case report

In January 2021, a female 1-year-old Kunekune was presented at the University Clinic for Swine with severe reduction of the field of vision resulting in prolonged reaction time when targeting barriers, due to moderate to severe thickening of the skin around both orbits also affecting the eyelids.Clinical examination revealed skin hyperplasia, nodular enlargement of the skin pores of the axillar and inguinal region. Ophthalmologists decided to remove parts of the thickened periocular skin, followed by histopathological examination.Once large amounts of demodectic mites were detected by histopathology, demodicosis could be diagnosed and treatment of the pig was started using sarolaner. Morphological and molecular analyses were performed. Histopathological and parasitological exams led to the aetiological diagnosis of demodicosis in the affected Kunekune pig. Severe skin lesions were revealed to be the consequence of an infestation with Demodex sp. Morphological analyses confirmed the involvement of D. phylloides. Molecular characterization indicated a Demodex species closely related to mites documented in wild boar - most probably D. phylloides for which no explicit sequences are available in GenBank yet. Treatment with sarolaner (2.6 mg/kg) resulted in a substantial regression of skin lesions, already detectable 1 month after first treatment.Demodicosis is a very rare disease in pigs that is most probably related to an impaired immune response to the mites. Demodectic mange should be included in the list of differential diagnoses in cases of periocular alterations of the skin of pigs

Effect of season and diet on heart rate and blood pressure in female red deer (Cervus elaphus) anaesthetised with medetomidine-tiletamine-zolazepam

Temperate zone ungulates like red deer (Cervus elaphus) show pronounced seasonal acclimatisation. Hypometabolism during winter is associated with cardiovascular changes, including a reduction in heart rate (fH) and temporal peripheral vasoconstriction. How anaesthesia with vasoactive substances such as medetomidine affect the seasonally acclimatised cardiovascular system is not yet known. We anaesthetised eleven healthy female red deer with medetomidine (0.1 mg/kg) and tiletamine/zolazepam (3 mg/kg) twice in winter (ad libitum and restricted feed) and in summer (ad libitum and restricted feed), with a two-week washout-period in-between, to test for the effect of season, food availability and supplementation with omega-3 or omega-6 polyunsaturated fatty acid (PUFA) on fH and arterial blood pressure (ABP) during anaesthesia. Six animals received pellets enriched with omega-6 fatty acids (FA), and five animals with omega-3 FA. Anaesthesia significantly decreased fH in summer but not in winter and ABP was lower in winter (p < 0.05). The combination of omega-6 FA enriched pellets and food restriction resulted in a lower fH and higher ABP during anaesthesia with more pronounced changes in winter (p < 0.001). Our results demonstrate that season, food availability and type of PUFA supplementation in red deer affect the cardiovascular system during anaesthesia

External mechanical perturbations challenge postural stability in dogs

This study aimed to explore the effect of external mechanical perturbations on postural stability (PS) in dogs using the body center of pressure (COP). Thirteen sound adult dogs were included in this study. PS was tested during quiet standing on a pressure measurement plate. The conditions included a standard standing measurement and external mechanical perturbations conducted using six settings on a motorized training platform with different intensities of speed and amplitude. Measurement conditions were compared using linear mixed-effects models, followed by multiple comparisons using Sidak\u27s alpha correction procedure. Compared with the standing measurement, external mechanical perturbations resulted in a significant increase in almost all COP parameters, indicating a challenge for the PS. Furthermore, an increase in amplitude had a greater effect than an increase in speed, whereas the combination of the highest intensities of amplitude and speed was not well tolerated by the dogs. The mediolateral COP displacement was significantly greater than the craniocaudal COP displacement during standing measurement and conditions with a small amplitude, whereas no significant difference was observed during settings with an increased amplitude. To the best of our knowledge, this is the first study to demonstrate the effects of a balance training device in dogs. Therefore, the intensity of the training programs on motorized platforms or similar devices can be controlled by the wobbling amplitude of the platform

Polarity reversal of canine intestinal organoids reduces proliferation and increases cell death

Apical-out intestinal organoids are a relatively simple method of gaining access to the apical cell surface and have faced increasing scientific interest over the last few years. Apical-out organoids can thus be used for disease modelling to compare differing effects on the basolateral versus the apical cell surface. However, these \u27inside-out\u27 organoids die relatively quickly and cannot be propagated as long as their basal-out counterparts. Here, we show that apical-out organoids have drastically reduced proliferative potential, as evidenced by immunohistochemical staining and the incorporation of the thymidine analogue EdU. At the same time, cell death levels are increased. Nevertheless, these phenomena cannot be explained by an induction of differentiation, as the gene expression of key marker genes for various cell types does not change over time

Tumor Cell Plasticity in Equine Papillomavirus-Positive Versus-Negative Squamous Cell Carcinoma of the Head and Neck

Squamous cell carcinoma of the head and neck (HNSCC) is a common malignant tumor in humans and animals. In humans, papillomavirus (PV)-induced HNSCCs have a better prognosis than papillomavirus-unrelated HNSCCs. The ability of tumor cells to switch from epithelial to mesenchymal, endothelial, or therapy-resistant stem-cell-like phenotypes promotes disease progression and metastasis. In equine HNSCC, PV-association and tumor cell phenotype switching are poorly understood. We screened 49 equine HNSCCs for equine PV (EcPV) type 2, 3 and 5 infection. Subsequently, PV-positive versus -negative lesions were analyzed for expression of selected epithelial (keratins, β-catenin), mesenchymal (vimentin), endothelial (COX-2), and stem-cell markers (CD271, CD44) by immunohistochemistry (IHC) and immunofluorescence (IF; keratins/vimentin, CD44/CD271 double-staining) to address tumor cell plasticity in relation to PV infection. Only EcPV2 PCR scored positive for 11/49 equine HNSCCs. IHC and IF from 11 EcPV2-positive and 11 EcPV2-negative tumors revealed epithelial-to-mesenchymal transition events, with vimentin-positive cells ranging between 50%. CD44- and CD271-staining disclosed the intralesional presence of infiltrative tumor cell fronts and double-positive tumor cell subsets independently of the PV infection status. Our findings are indicative of (partial) epithelial-mesenchymal transition events giving rise to hybrid epithelial/mesenchymal and stem-cell-like tumor cell phenotypes in equine HNSCCs and suggest CD44 and CD271 as potential malignancy markers that merit to be further explored in the horse

Influence of sperm cryopreservation on sperm motility and proAKAP4 concentration in mice

The protein proAKAP4 is crucial for sperm motility and has been suggested as an indicator of male fertility. We determined the relationship between proAKAP4 concentration and sperm motility parameters in mice, and investigated the effects of cryopreservation on these variables.Computer-assisted sperm analysis and ELISA were applied to determine sperm motility and proAKAP4 concentration in fresh and frozen-thawed epididymal sperm of SWISS, B6D2F1, C57BL/6N, and BALB/c mice.ProAKAP4 levels ranged between 12 and 89 ng/ml and did not differ between fresh and frozen-thawed samples, or between strains. We found a negative relationship between proAKAP4 levels and some sperm motility parameters. Sperm traits differed between strains, and cryopreservation negatively affected sperm velocity but not sperm direction parameters.ProAKAP4 levels in epididymal mouse spermatozoa were unaffected by cryopreservation, highlighting the robustness of this parameter as a potentially time-independent marker for sperm motility and fertility. The high individual variation in proAKAP4 levels supports the potential role of proAKAP4 as a marker for sperm quality, though we found no positive, and even negative relationships between proAKAP4 levels and some sperm motility parameters. Future studies have to investigate the significance of proAKAP4 as an indicator for fertility in mice

Risk factors associated with post-weaning diarrhoea in Austrian piglet-producing farms

Post-weaning diarrhoea (PWD) is a frequent, multifactorial disease of piglets leading to increased mortality rates and high economic losses. Due to the emergence of multi-resistant Escherichia coli isolates and the ban of zinc oxide (ZnO) in the EU since June 2022, alternative measures to prevent PWD are urgently needed. While an abundance of feed supplements is described to prevent PWD, there are hardly any studies reflecting the current situation of PWD in the field. Thus, we aimed to identify differences in management practices, housing and feeding strategies between farms with PWD and farms without PWD. Data were personally collected using a semi-structured questionnaire in 257 Austrian piglet-producing farms. Farms with PWD in more than 10% of all weaned groups within twelve months prior to data collection were defined as case farms (n = 101), while the remaining 136 farms were defined as control farms. Data from 237 farms and 69 explanatory variables were analysed via penalized binary logistic regression using elastic-net in 100 different splits into randomly selected training and test datasets (80:20). Treatment with ZnO and/or colistin (136 farms) was negatively associated with PWD in all splits and had the biggest estimated absolute log odds ratio out of all tested variables. Implementation of an all-in/all-out system in the nursery units and administration of probiotics or horseradish also had preventive effects in most splits (≥ 97%). A higher number of feeding phases for piglets within the first seven weeks of life and housing on fully slatted floors was associated negatively with the occurrence of PWD as well in > 95% of all splits. PWD was more likely to occur on farms having problems with neonatal diarrhoea or postpartum dysgalactia syndrome. While our data demonstrate that treatment with ZnO or colistin had the biggest statistical effect on PWD, we were able to identify other preventive measures like supplementation with probiotics or horseradish. Since implementation of all-in/all-out measures and fully slatted floors were also negatively associated with the occurrence of PWD on visited farms, we assume that reduction of bacterial load by the implementation of simple hygiene measures are still crucial to prevent PWD

Dynamics and Diversity of Intrauterine Anaerobic Microbiota in Dairy Cows with Clinical and Subclinical Endometritis

The aim of the study was to characterize the dynamics of anaerobic cultivable postpartum microbiota in the uterus of dairy cows. In total, 122 dairy cows were enrolled and sampled on day 0 (day of calving) and on days 3, 9, 15, 21, and 28 postpartum (pp). Samples were cultivated anaerobically and analyzed by MALDI-TOF MS. In total, 1858 isolates were recovered. The most prevalent facultative anaerobic genera were Trueperella (27.8%), Streptococcus (25.4%), and Escherichia (13.1%). The most prevalent obligate anaerobes were Peptoniphilus (9.3%), Bacteroides (3.3%), and Clostridium (2.4%). The microbial communities were highly dynamic and diverse. On the animal level, Trueperella pyogenes on day 21 and 28 pp was associated with clinical endometritis, and E. coli on day 21 pp was associated with subclinical endometritis. The occurrence of Streptococcus pluranimalium on day 28 was related to uterine health. The presence of T. pyogenes, Streptococcus, and Peptoniphilus was significantly associated with an increased risk for purulent vaginal discharge. Primiparous cows showed a higher prevalence of T. pyogenes, Fusobacterium necrophorum, Porphyromonas levii, and Peptoniphilus spp. than multiparous cows but were not more susceptible to uterine diseases. This study might provide a suitable basis for future co-cultivation studies to elucidate potential synergistic interactions between microbiota

Haemonchosis in Sheep and Goats, Control Strategies and Development of Vaccines against Haemonchus contortus

The evolutionary success of parasitic worms causes significant economic losses and animal health problems, including in the small ruminant industry. The hematophagous nematode Haemonchus contortus is a common endoparasite that infects wild and domestic ruminants worldwide, especially in tropical and subtropical regions. To date, the most commonly applied control strategy is the administration of anthelminthic drugs. The main disadvantages of these chemicals are their ecotoxic effects, the necessary withdrawal period (especially important in dairy animals) and the increasing development of resistance. Vaccines offer an attractive alternative control strategy against Haemonchus infections. In previous years, several potential vaccine antigens prepared from H. contortus using the latest technologies have been assessed in clinical trials using different methods and strategies. This review highlights the current state of knowledge on anti-H. contortus vaccines (covering native, recombinant and DNA-based vaccines), including an evaluation, as well a discussion of the challenges and achievements in developing protective, efficient, and long-lasting vaccines to control H. contortus infection and haemonchosis in small ruminants. This paper also addresses novel developments tackling the challenge of glycosylation of putative candidates in recombinant form