Inhibition of SARS-CoV-2 by type I and type III interferons.

The recently emerged severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is the causative agent of the devastating COVID-19 lung disease pandemic. Here, we tested the inhibitory activities of the antiviral interferons of type I (IFN-α) and type III (IFN-λ) against SARS-CoV-2 and compared them with those against SARS-CoV-1, which emerged in 2003. Using two mammalian epithelial cell lines (human Calu-3 and simian Vero E6), we found that both IFNs dose-dependently inhibit SARS-CoV-2. In contrast, SARS-CoV-1 was restricted only by IFN-α in these cell lines. SARS-CoV-2 generally exhibited a broader IFN sensitivity than SARS-CoV-1. Moreover, ruxolitinib, an inhibitor of IFN-triggered Janus kinase/signal transducer and activator of transcription signaling, boosted SARS-CoV-2 replication in the IFN-competent Calu-3 cells. We conclude that SARS-CoV-2 is sensitive to exogenously added IFNs. This finding suggests that type I and especially the less adverse effect-prone type III IFN are good candidates for the management of COVID-19.

Downregulation of testicular function in the goat by altrenogest.

BACKGROUND: The present study investigated whether the administration of the progestin altrenogest provides noninvasive, temporary, and reversible suppression of gonadal function in the goat as a potential alternative to chirurgical castration, which is related with irreversibility, risks of complications till death of the animal and welfare issues. Eight sexually mature Peacock goats were randomly divided into two groups. The experimental group was administered altrenogest (0.088 mg/kg) orally once daily for 7 weeks. The remaining four goats received an oral glucose solution and served as the control group. After completing the administration period, the reversibility of the medication was evaluated for another 7 weeks (observation phase). The treatment effects were assessed by clinical examination; ultrasound examination of the testes, including one-dimensional grayscale analysis, blood testosterone levels, analysis of semen parameters and libido. At the end of the observation period, the animals were castrated and the testicles were examined histologically. RESULTS: Altrenogest treatment had no significant effect on the physical development of the goats, the sonographic appearance of the testes, the gray values measured in the ultrasound images, or the blood testosterone levels. The effects of treatment on the testicular and semen parameters varied widely in the experimental animals; the testicle volume was significantly lower and the number of pathologically altered sperm in the ejaculate was significantly higher in treated animals. CONCLUSION: These findings indicate that daily altrenogest administration at a dose of 0.088 mg/kg does not reliably suppress gonadal function in the goat.

Associations between the metabolic status of the cow and colostrum quality as determined by Brix refractometry.

Supplying newborn calves with immunoglobins is critical for their health and a daily challenge in the dairy industry. Among various factors determining colostrum quality, the prepartum metabolic status of the cow might be of particular importance. The objective of this observational cross-sectional study was to evaluate relationships between cow-level variables and the colostrum quality as determined by Brix refractometry. A total of 873 cows of varying breed and parity from 124 German dairy herds were included in the study, and blood and urine samples were taken 3 to 1 wk before the expected calving date. Effectively, samples were collected on average 8.2 d (geometric mean) before calving, ranging from 2 to 45 d. The final variable set included body condition score, lameness score, breed, parity, vaccination of the cow, the activity of glutamate dehydrogenase and aspartate aminotransferase, the urine concentration of creatinine, net acid-base excretion, the serum concentration of cholesterol and calcium, and the difference in albumin and total protein concentration. Generalized linear mixed effects regression models with hierarchically structured random effects (cow within herd) using the maximum likelihood method were fitted to the data to identify associations between the Brix value as an outcome and cow-level variables as predictors. Cows entering second parity had lower Brix values compared with cows entering third or greater parity, and prepartum vaccination of cows led to higher Brix values compared with nonvaccinated cows. Cows with a moderate to high lameness score had lower Brix values than cows with low-grade lameness. An increase of glutamate dehydrogenase serum activity and serum calcium concentration were associated with lower Brix values, whereas an increase in the difference of total protein and albumin serum concentration led to higher Brix values. In conclusion, the metabolic health of the cow affects colostrum quality and may cause failure of passive immunoglobulin transfer as well as impaired calf health.

Serosurvey of Treponema pallidum infection among children with skin ulcers in the Tarangire-Manyara ecosystem, northern Tanzania.

BACKGROUND: The first yaws eradication campaign reduced the prevalence of yaws by 95%. In recent years, however, yaws has reemerged and is currently subject to a second, ongoing eradication campaign. Yet, the epidemiological status of Tanzania and 75 other countries with a known history of human yaws is currently unknown. Contrary to the situation in humans in Tanzania, recent infection of nonhuman primates (NHPs) with the yaws bacterium Treponema pallidum subsp. pertenue (TPE) have been reported. In this study, we consider a One Health approach to investigate yaws and describe skin ulcers and corresponding T. pallidum serology results among children living in the Tarangire-Manyara ecosystem, an area with increasing wildlife-human interaction in northern Tanzania. METHODS: To investigate human yaws in Tanzania, we conducted a cross-sectional study to screen and interview skin-ulcerated children aged 6 to 15 years, who live in close proximity to two national parks with high numbers of naturally TPE-infected monkeys. Serum samples from children with skin ulcers were tested for antibodies against the bacterium using a treponemal (Treponema pallidum Particle Agglutination assay) and a non-treponemal (Rapid Plasma Reagin) test. RESULTS: A total of 186 children aged between 6 and 15 years (boys: 10.7 ± 2.1 (mean ± SD), N = 132; girls: 10.9 ± 2.0 (mean ± SD), N = 54) were enrolled. Seven children were sampled at health care facilities and 179 at primary schools. 38 children (20.4%) reported active participation in bushmeat hunting and consumption and 26 (13.9%) reported at least one physical contact with a NHP. None of the lesions seen were pathognomonic for yaws. Two children tested positive for treponemal antibodies (1.2%) in the treponemal test, but remained negative in the non-treponemal test. CONCLUSIONS: We found no serological evidence of yaws among children in the Tarangire-Manyara ecosystem. Nevertheless, the close genetic relationship of human and NHPs infecting TPE strains should lead to contact prevention with infected NHPs. Further research investigations are warranted to study the causes and possible prevention measures of spontaneous chronic ulcers among children in rural Tanzania and to certify that the country is free from human yaws.

Effect of heel elevation on breakover phase in horses with laminitis.

BACKGROUND: In a laminitic horse, the maximal loading of the toe region occurs during the breakover phase. To date, no kinetic data demonstrates the effect of supportive orthopaedic therapy in horses with laminitis on breakover phase. Thus, the purpose of this study was to examine the effect of heel elevation on the breakover phase. Eight horses with acute laminitis treated medically as well as with application of a hoof cast with heel wedge (HCHW) were included in this study. Immediately following cessation of clinical signs of acute laminitis, two measurements using the Hoof™ System were taken: the first with HCHW and the second immediately following removal of the HCHW, i.e. in barefoot condition (BFC). The hoof print was divided into three regions: toe, middle hoof, and heel. Kinetic parameters included vertical force (VF), stance duration, contact area (CA) for all hoof regions during stance phase, duration of breakover, VF in the toe region at onset of breakover and location of centre of force. RESULTS: The VF and CA were higher in the heel region (63 and 61%, respectively) and decreased significantly after removal of the HCHW (43 and 28% after removal, respectively). The breakover phase in horses with HCHW lasted 2% of stance phase and was significantly shorter than that in BFC, which lasted 6% of stance phase. The VF at onset of breakover for the toe region in horses with HCHW was significantly lower than that in BFC. The centre of the force was located at the heel region in all horses with the HCHW, and at the middle the hoof region in BFC. CONCLUSIONS: Heel elevation in horses with laminitis as examined on a concrete surface significantly shortens breakover phase and decreases the vertical force in the toe region during breakover. HCHW provides adequate support to the palmar hoof structures by increasing the contact area in the heel region and incorporating the palmar part of frog and sole into weight bearing, thus decreasing the stress on the lamellae. Hoof cast with heel elevation could be a beneficial orthopaedic supportive therapy for horses suffering from acute laminitis.

Viability assessment of spermatozoa in large falcons (Falco spp.) using various staining protocols.

Viability assessment is an important part of semen analysis, and various live/dead staining protocols have been used in semen of avian species. Results of live/dead count differed between dyes, staining protocols and bird species, impeding comparability between studies and requiring species-specific comparisons of viability stains. In raptor semen, similar comparisons are absent. Thus, the aim of the present study was to compare eight conventional viability stains. Eosin blue 2% [EB], eosin blue 2% with the addition of 3% sodium citrate [EB2], eosin blue-nigrosin 5% [EBN5], eosin yellow-nigrosin 5% [EYN5], eosin yellow-nigrosin 10% [EYN10], eosin blue-aniline blue [EBA], eosin yellow-aniline blue [EYA] and bromophenol blue-nigrosin [BBN] were evaluated in comparison with the fluorescence stain SYBR® Green-propidium iodide [SYBR-PI] in spermatozoa of falcons. The comparison was performed using conventional light microscopy which is applicable in breeding centres, veterinary practices and field studies. Additionally, live/dead stains were correlated to motility values of the same samples to validate sperm viability. Light microscopy using EB and using SYBR-PI enabled an effective and clear differentiation between alive and dead spermatozoa of falcons. Motility values correlated significantly and strongly with EB only (r = .629; p < .001), but not with any other stain used in the study. Therefore, our results suggest EB as the most suitable stain for viability assessment in the semen of large falcons.

Interaction of different Chlamydiae species with bovine spermatozoa.

BACKGROUND: Interaction of spermatozoa and Chlamydiae spp. might contribute to reduced fertility in cattle. To proof this hypothesis, bovine semen was incubated with viable or heat inactivated Chlamydia (C.) abortus or psittaci (Multiplicity of infection = 1) and sperm motility was monitored with a computer-assisted sperm analyzer over 24 h. Additionally, the interaction with the spermatozoa was further investigated by means of light and transmission electron microscopy (TEM). RESULTS: Only viable Chlamydiae of both species decreased sperm motility and this only after about 9 h. Taking binding rates into account, the loss of sperm motility after about 9 h could likely be a consequence of Chlamydiae attachment to the spermatozoa. About two thirds of the Chlamydiae elementary bodies were bound to the front third of the sperm, the acrosomal region. No inclusions of Chlamydiae in spermatozoa were observed in TEM after 2 h co-incubation. CONCLUSIONS: As initial motility was not affected following co-incubation of viable Chlamydiae and bovine sperm, it seems likely that sperm could serve as a carrier/vehicle for Chlamydiae facilitating cervical passage of Chlamydiae spp. in cattle. Additionally, our results suggest that spermatozoa carrying Chlamydiae may have no initial disadvantage in reaching the oviduct, but are immotile at the time of ovulation what might have an impact on fertilization capacities of the individual sperm. Consequently, high concentrations of the investigated Chlamydiae in the seminal plasma or female genital tract might play a role in reduced fertility in cattle.

Optimization of RNA extraction protocol for long-term archived formalin-fixed paraffin-embedded tissues of horses.

A suitable RNA extraction protocol was established to gain high quality RNA from formalin-fixed paraffin-embedded tissues to perform reliable molecular assays either applicable for using FFPE tissue archives or tissues with harsh formalin-fixation. Eighteen FFPE samples from the central nervous system of horses, stored up to 11 years, were used as archive cases. To test the influence of the fixation period, brain, liver, kidney, and skeletal muscle tissue fragments from another horse, were treated either with water or tris-acetate-EDTA buffer after fixation under different timepoints with 10% unbuffered formalin. Two deparaffinization methods and three proteinase K-based lysis step were tested and translated into three protocols. After detailed statistical analysis it was determined that a longer period and increase in volume of proteinase K incubation provide higher yields and purity of RNA (P < 0.01) of archived samples. Alongside, amplification of equid-housekeeping gene up to 298 bp was successful with the protocol adaptations. For different formalin-fixation timepoints, it was demonstrated that the right choice for treatment and formalin-fixation period is organ-related (P ≤ 0.05). Essentially, little alterations to pre-existing extraction protocols unwound the RNA of up to 11-year-old samples, enabling the use of FFPE tissue archives or e.g. harshly fixed material needed in infection research under high biosafety levels for a variety of molecular analysis.

Effect of temperature and time after collection on buck sperm quality.

BACKGROUND: Different parameters are assessed as part of the semen analysis but a standard protocol for evaluation of goat semen is still missing. The aim of this study was to analyse two different factors affecting buck sperm quality in the post-collection period prior to adding the extender. Here we examined the effects of two handling temperatures (20 °C, 37 °C) and various examination time points (3-30 min) after semen collection. RESULTS: Examination time point had a significant influence on raw sperm viability (p < 0.05), motility (p < 0.05) and on semen pH (p < 0.05). The two different handling temperatures had no significant effect on sperm viability (p > 0.05), motility (p > 0.05), with the exception of fast moving sperm (p = 0.04), or on semen pH (p > 0.05). CONCLUSION: Examination time point was identified as factor strongly influencing raw peacock buck semen after collection. Raw goat semen can tolerate room temperatures for at least 10 min without impacting overall semen quality. In order to obtain comparable results, semen samples should always be examined within 10 min after collection.

Sensitivity and specificity of magnetic resonance imaging and computed tomography for the determination of the developmental state of cranial sutures and synchondroses in the dog.

BACKGROUND: During skull ontogenesis, growth centers in the skull base and calvarial bones allow gradual expansion of the cranial vault. Premature growth termination of cranial base synchondroses and/or calvarial sutures can result in devastating skull dysmorphologies. There is evidence to believe that a premature closure in one or more cranial growth centers contribute to the brachycephalic skull morphology in dogs. To provide a proof of concept for the non-invasive investigation of ontogenetic changes in cranial sutures and synchondroses in living dogs, we compared magnet resonance imaging (MRI) and computed tomography (CT) with histologic findings. Our aim was to determine the in vitro sensitivity and specificity for conventional clinical imaging methods in the assessment of cranial suture closure and synchondroses ossification in dogs. RESULTS: The evaluation of cranial base synchondroses in MRI had a sensitivity of up to 93.1% and a specificity of 72.7% dependent on the observer. The evaluation of cranial base synchondroses in CT had a sensitivity of 92.2% and a specificity of 86.4%. Suture assessment on MRI suture assessment had a sensitivity of 82.1% dependent on the observer and a specificity of 19.3%. CT suture assessment had a sensitivity of 85.1% and a specificity of 40.4% in dependence of the observer. CONCLUSION: Conventional cross-sectional imaging techniques (MRI and CT) allow reliable assessment of the open or closed state of synchondroses within the cranial base. In contrast CT and MRI are not suitable for a reliable assessment of the cranial sutures in dogs.

Concentrations of minerals in the canine prostatic fluid.

Current knowledge about the composition of the prostate fluid in healthy male dogs is limited and restricted to small case numbers. Furthermore, published data often vary significantly regarding sample processing and analytical methods. Therefore, we aimed to provide data on the composition of electrolytes and minerals in the canine prostatic fluid in a larger population (n = 30 dogs/samples) and to compare these results with the existing literature. Concentrations of sodium, potassium and copper analysed in our population were most consistent with those in the literature. Different to this, concentrations of total calcium, magnesium, zinc and inorganic phosphate varied. Whereas magnesium, zinc and inorganic phosphate seemed to depend on the analysis method, total calcium concentrations differed if centrifugation was performed or not. Our results clearly indicate a need for standardization of methods for analysis of seminal plasma components.

Single tracheal inoculation of Aspergillus fumigatus conidia induced aspergillosis in juvenile falcons (Falco spp.).

Aspergillosis is a common and life-threatening respiratory disease in raptors with acute and chronic courses. Among raptors, gyrfalcons (Falco rusticolus) and their hybrids are often declared to be highly susceptible with juvenile individuals being the most susceptible. However, species- and age-specific experimental studies are lacking and minimal infective doses (IDs) for Aspergillus spp. conidia are unknown.Therefore, 8-week-old, healthy gyr-hybrid falcons (F. rusticolus X F. cherrug) (N = 18) were experimentally infected with Aspergillus fumigatus using a single intratracheal inoculation with varying dosages of conidia (102 to 107 conidia). Over 28 days, clinical signs were monitored as well as haematological and serological parameters. Following euthanasia, necropsy, histopathology, bacteriology, and mycology were performed. Re-isolated fungi were compared to the inoculum using microsatellite length polymorphisms. During the trial, clinical signs and dyspnoea correlated significantly with the ID. Necropsy revealed fungal lesions in the upper and lower airways of 10/18 inoculated falcons, but not in the control birds. In 9/18 inoculated falcons, fungal granulomas were confirmed in histopathology and A. fumigatus was re-isolated from these granulomas. Except one nasal isolate all re-isolated fungal strains were identical to the inoculum strain. Based on mycology and histopathology a minimal ID of 50% was calculated to be MID50% (±S.E.) = 104.52±0.67 for a single tracheal inoculation of A. fumigatus conidia. This study demonstrates for the first time that a single exposure is able to cause acute aspergillosis in juvenile falcons.

Canine calcium oxalate urolithiasis: Frequency of Whewellite and Weddellite stones from 1979 to 2015.

This study reports on a retrospective evaluation of epidemiological data from calcium oxalate stones in dogs differentiated into calcium oxalate monohydrate (Whewellite, Wh) and calcium oxalate dihydrate (Weddellite, Wd). Of the 22 456 uroliths submitted from 1979 to 2015, 6690 (29.8%) were composed of > 70% calcium oxalate. During the observation period, the proportion of calcium oxalate stones rose from 4% (1979) to 46% (2015). Of all the calcium oxalate stones, 31.0% were Wh and 49.4% Wd, while 19.6% were a mixture of Wh and Wd. The dogs with Wh stones were significantly older than the dogs with Wd stones. Several breeds have increased odds ratios (OR) for either Wh (5 highest OR: Norwich terrier, keeshond, Norfolk terrier, fox terrier, sheltie) or Wd (Pomeranian, borzoi, Japanese spitz, Finnish lapphund, bichon frise). Analytical differentiation of the calcium oxalate stones into Wh and Wd is important for understanding the cause and possible treatment and prevention of the uroliths.

Urolithiase d'oxalate de calcium chez les chiens : fréquence des calculs Whewellite et Weddellite de 1979 à 2015. Cette étude présente un rapport sur l'évaluation rétrospective de données épidémiologiques de calculs d'oxalate de calcium chez les chiens qui ont été différenciés en monohydrate d'oxalate de calcium (Whewellite, Wh) et en dihydrate d'oxalate de calcium (Weddellite, Wd). Parmi les 22 456 urolithes soumis de 1979 à 2015, 6690 (29,8 %) étaient composés de > 70 % d'oxalate de calcium. Durant la période d'observation, la proportion des calculs d'oxalate de calcium est passée de 4 % (1979) à 46 % (2015). Parmi tous les calculs d'oxalate de calcium, 31,0 % étaient Wh et 49,4 % étaient Wd, tandis que 19,6 % étaient un mélange de Wh et de Wd. Les chiens avec des calculs Wh étaient significativement plus âgés que les chiens ayant des calculs Wd. Plusieurs races présentent des ratios de probabilité supérieure (RP) pour soit Wh (5 RP plus élevés : Terrier de Norwich terrier, Keeshond, Terrier de Norfolk, Fox terrier, Shetland) ou Wd (Poméranien, Barzoï, Spitz japonais, Finnois de Laponie, Bichon frisé). La différenciation analytique des calculs d'oxalate de calcium selon Wh et Wd est importante pour comprendre la cause ainsi que pour le traitement et la prévention éventuelle des urolithes.(Traduit par Isabelle Vallières).

Start early! Does social instability during the pre- and early postnatal development prepare male wild cavies for social challenge later in life?

BACKGROUND: The social environment the mother experiences during pregnancy and lactation can powerfully influence the offspring's behavioural profile. Our previous studies in wild cavies show that two different social environments during pregnancy and lactation bring about different behavioural strategies of male offspring later in life: An unstable social environment leads to a behavioural camouflage strategy, hypothesised to be beneficial at times of socially challenging situations. A stable social environment during early phases of life, however, leads to an early reproduction strategy, expected to be more successful at times of social stability. In the present study, we observed the behavioural strategies of the two types of males in direct comparison in a socially challenging situation: Two adolescent males were placed simultaneously in an unknown social group consisting of one adult male and two females in a semi-naturalistic environment. Cortisol as well as testosterone concentrations and activity levels were compared. Furthermore, paternities were analysed after the males reached sexual maturity. We hypothesised that sons showing a behavioural camouflage strategy are better adapted to cope with this socially challenging situation compared to those displaying an early reproduction strategy. RESULTS: At the beginning of the experiment, no differences in plasma cortisol concentrations between the males were found, both showed a highly significant increase due to the challenging situation. From day 5 until the end of the experiment (duration = 40 days) sons showing an early reproduction strategy had significantly higher plasma cortisol concentrations compared with those showing a behavioural camouflage strategy. Plasma testosterone concentrations did not differ significantly. Activity levels decreased significantly over time independently of the male's behavioural strategy. Both types of males did not sire offspring during the observation period. CONCLUSION: Higher cortisol values from day 5 until the end of the experiment in sons showing an early reproduction strategy indicate higher levels of stress in these males compared to those displaying a camouflage strategy. We conclude that the modulation of the males behavioural strategy due to an unstable social environment during early development facilitates the endocrine adaptation to a comparable social situation later in life.

Diffusion-weighted imaging of the brains of dogs with idiopathic epilepsy.

BACKGROUND: Idiopathic epilepsy is one of the most common neurological disorders in dogs. Unfortunately, up to 30% of dogs with idiopathic epilepsy show no improvement under antiepileptic drug treatment. Diffusion-weighted imaging is used in human medicine to identify epileptogenic foci in the brain to allow for more invasive treatments such as deep brain stimulation or surgical removal. The aim of this study was to ass the feasibility of interictal diffusion-weighted MRI in dogs and to evaluate the distribution of diffusion in the brains of dogs with idiopathic epilepsy (IE) and to compare these values to previously published values from healthy beagle dogs. Client-owned dogs with the final diagnosis of IE were included in this study. MRI examination was carried out using a 1.0Tesla superconductive magnet. Diffusion-weighted images using a single shot echo planar imaging sequence (SSh-EPI) with a b value of b = 0 s/mm2 and b = 800 s/mm2 were acquired in a dorsal and transverse plane with diffusion gradients in all three planes (x-, y- and z-plane). An ADC (apparent diffusion coefficient) map of the isometric image of each acquired slice was generated. Regions of interest (ROIs) were manually drawn around the caudate nucleus, the thalamus, the piriform lobe including the amygdala, the hippocampus, the semioval center and the temporal cerebral cortex by one of the authors. ROI drawings were repeated 5 times at different time points to assess intra-obersver variability. A multi-way mixed-model analysis of variance (ANOVA) and two-way ANOVA were used during statistical analysis. A p value of p < 0.05 was considered significant. RESULTS: Dogs with IE showed a significantly increased ADC in the amygdala within the piriform lobe and in the semioval center (p < 0.05) compared with the healthy control group. CONCLUSION: Changes in the piriform lobe in cases of epilepsy are reported infrequently in human and veterinary medicine. Similar to our results, ADC changes in the interictal phase usually include an increase in ADC due to cell loss and increased intercellular spaces. Diffusion MRI might be a promising technique for the examination of canine epileptic patients lacking other gross neuromorphological abnormalities.

Evaluation of different diagnostic methods for the detection of Mycobacterium avium subsp. paratuberculosis in boot swabs and liquid manure samples.

BACKGROUND: Environmental sampling based on boot swabs and/or liquid manure samples is an upcoming strategy for the identification of paratuberculosis (paraTB) positive herds, but only limited data are available regarding the diagnostic performance of molecular detection methods (qPCR) versus faecal culture (FC) for this purpose. In the present study, the test characteristics of two different qPCR protocols (A and B) and a standardized FC protocol, for the detection of Mycobacterium avium subsp. paratuberculosis in boot swabs and liquid manure samples were evaluated. RESULTS: In 19 paraTB unsuspicious and 58 paraTB positive herds boot swabs and liquid manure were sampled simultaneously and analyzed in three different diagnostic laboratories. Using boot swabs and liquid manure, a substantial to excellent accordance was found between both qPCRs, for boot swabs also with culture, while for liquid manure the detection rate of culture was decreased after prolonged storage at -20 °C. The quantitative results of both qPCR methods correlated well for the same sample and also for boot swabs and liquid manure from the same herd. When cut-off threshold cycle (CT-)-values were applied as recommended by the manufacturers, herd level specificity (Sp) of qPCR B was below 100% for boot swabs and for both qPCRs for liquid manure. A decreased herd level sensitivity was encountered after adjustment of Sp to 100% and re-calculation of the cut-off CT-values. CONCLUSIONS: qPCR is equally suitable as bacterial culture for the detection of Mycobacterium avium subsp. paratuberculosis in boot swabs and liquid manure samples. Both matrices represent easily accessible composite environmental samples which can be tested with reliable results. The data encourage qPCR testing of composite environmental samples for paraTB herd diagnosis.

Behaviour of adipose-derived canine mesenchymal stem cells after superparamagnetic iron oxide nanoparticles labelling for magnetic resonance imaging.

BACKGROUND: Therapy with mesenchymal stem cells (MSCs) has been reported to provide beneficial effects in the treatment of neurological and orthopaedic disorders in dogs. The exact mechanism of action is poorly understood. Magnetic resonance imaging (MRI) gives the opportunity to observe MSCs after clinical administration. To visualise MSCs with the help of MRI, labelling with an MRI contrast agent is necessary. However, it must be clarified whether there is any negative influence on cell function and viability after labelling prior to clinical administration. RESULTS: For the purpose of the study, seven samples with canine adipose-derived stem cells were incubated with superparamagnetic iron oxide nanoparticles (SPIO: 319.2 µg/mL Fe) for 24 h. The internalisation of the iron particles occurred via endocytosis. SPIO particles were localized as free clusters in the cytoplasm or within lysosomes depending on the time of investigation. The efficiency of the labelling was investigated using Prussian blue staining and MACS assay. After 3 weeks the percentage of SPIO labelled canine stem cells decreased. Phalloidin staining showed no negative effect on the cytoskeleton. Labelled cells underwent osteogenic and adipogenic differentiation. Chondrogenic differentiation occurred to a lesser extent compared with a control sample. MTT-Test and wound healing assay showed no influence of labelling on the proliferation. The duration of SPIO labelling was assessed using a 1 Tesla clinical MRI scanner and T2 weighted turbo spin echo and T2 weighted gradient echo MRI sequences 1, 2 and 3 weeks after labelling. The hypointensity caused by SPIO lasted for 3 weeks in both sequences. CONCLUSIONS: An Endorem labelling concentration of 319.2 µg/mL Fe (448 µg/mL SPIO) had no adverse effects on the viability of canine ASCs. Therefore, this contrast agent could be used as a model for iron oxide labelling agents. However, the tracking ability in vivo has to be evaluated in further studies.

Occlusal Angles of Equine Incisors.

The angulation of equine incisors is frequently used as a parameter for dental corrections. However, the term incisor angle is only vaguely defined, and no studies exist presenting a series of reliable measurements in individual incisors of multiple horses. The aim of this study was to establish an exact method to determine incisor angles and to test whether clinically accessible landmarks (facial crest and bars) are suitable to estimate incisor angles. Eighteen horses were used to create 3-dimensional (3D) reconstructions of the skulls from computed tomography (CT) data sets. Reference planes (median and transverse plane) were calculated using defined anatomical landmarks. Subsequently, occlusal planes for incisors and for incisor quadrants were calculated. Occlusal table angles were measured in relation to the reference planes. For each incisor, sagittal and transverse angles were measured. Mean values of individual incisor angles ranged from 3.5° to 6.8° (transverse angle) and from 32.6° to 44.9° (sagittal angle). No significant differences were detected in mean between the left and the right side when teeth in same Triadan positions were compared. However, in individual horses, marked differences between the left and the right side of the jaws occurred. Mandibular incisors showed significantly steeper sagittal angles than maxillary incisors. Furthermore, angles of opposing incisors were correlated with each other. The facial crest and the upper jaw bars featured a curved shape and were therefore of limited use to estimate the angulation of the upper incisors. In contrast, the lower jaw bars were suitable to determine the angulations of lower incisors.

Perfluoroalkyl Acid Concentrations in Livers of Fox (Vulpes vulpes) and Chamois (Rupicapra rupicapra) from Germany and Austria.

The concentrations of 11 perfluorinated alkyl acids (PFAAs) were measured in the livers of foxes (Vulpes vulpes) from Germany, a primarily carnivorous species, and chamois (Rupicapra rupicapra) from Austria, an herbivorous species. Perfluorooctanesulfonate (PFOS) at concentrations [all results refer to wet weight (ww)] of 3.2-320 µg/kg were detected in all 40 fox livers tested, yielding an arithmetic mean of 46.6 µg/kg and a median of 29.8 µg/kg. Long-chain PFAAs were detected at concentrations of 1.7 µg/kg perfluorononanoic acid (PFNA) to 2.4 µg/kg perfluorodecanoic acid (PFDA) and perfluorododecanoic acid (PFDoDA). Of the short-chain PFAAs tested, only perfluorohexanoic acid (PFHxA) was found in 1 fox liver at a concentration of 1.4 µg/kg, and perfluorohexane sulfonate (PFHxS) was found in 2 fox livers at a concentration of 1 µg/kg each. PFOS and PFNA concentrations higher than limit of quantification (LOQ) were detected in 90.9 and 81.8 % of chamois livers, respectively. The arithmetic mean for PFOS concentrations was 2.2 µg/kg (median 2.4 µg/kg), a factor of 21 (median factor of 12) lower than in fox livers. The arithmetic mean for PFNA concentrations was 2.0 µg/kg (median 1.9 µg/kg). Perfluorobutanoic acid, PFHxA, perfluorooctanoic acid, perfluorobutanesulfonate, and PFHxS were not detected at concentrations higher than the LOQ in any of the samples. The various results are compared with one another and with the results of other studies of herbivorous, carnivorous, and omnivorous wild animals. The highest concentrations of PFAA, in particular PFOS, were found in omnivorous animals followed by carnivores. The lowest levels were present in herbivores.

Magnetic resonance imaging signs of high intraventricular pressure--comparison of findings in dogs with clinically relevant internal hydrocephalus and asymptomatic dogs with ventriculomegaly.

BACKGROUND: Magnetic resonance imaging (MRI) findings of canine brains with enlarged ventricles in asymptomatic dogs were compared to those in dogs with clinically relevant internal hydrocephalus, in order to determine the imaging findings indicative of a relevant increase in intraventricular pressure. Discrimination between clinically relevant hydrocephalus and ventriculomegaly based on MRI findings has not been established yet and is anything but trivial because of the wide variation in ventricular size in different dog breeds and individuals. The MRI scans of the brains of 67 dogs of various breeds, skull conformation and weight were reviewed retrospectively. Based on clinical and imaging findings, the dogs were divided into three groups: a normal group (n = 20), a group with clinically silent ventriculomegaly (n = 25) and a group with severe clinically relevant internal hydrocephalus (n = 22). In addition to the ventricle/brain-index, a number of potential subjective signs of increased intraventricular pressure were recorded and compared between the groups. RESULTS: The ventricle/brain-index was significantly higher in dogs with relevant hydrocephalus (p < 0.001) and a threshold value of 0.6 was specified as a discriminator between internal hydrocephalus and ventriculomegaly. Other MR imaging findings associated with clinically relevant hydrocephalus were an elevation of the corpus callosum (p < 0.01), dorsoventral flattening of the interthalamic adhesion (p < 0.0001), periventricular edema (p < 0.0001), dilation of the olfactory recesses (p < 0.0001), thinning of the cortical sulci (p < 0.0001) and/or the subarachnoid space (p < 0.0027) and disruption of the internal capsule adjacent to the caudate nucleus (p < 0.0001). CONCLUSION: A combination of the abovementioned criteria may support a diagnosis of hydrocephalus that requires treatment.