Carp oedema virus disease outbreaks in Czech and Slovak aquaculture

This work describes the first confirmed cases of carp oedema virus disease (CEVD) in Slovakia and the Czech Republic and the phylogenetic analysis of Czech and Slovak carp oedema virus (CEV) isolates. Four cases of disease outbreak in the Czech Republic are described, the oldest dating from mid-May 2013 and one case from Slovakia dating from May 2019. In all cases, virus presence was confirmed using nested PCR. PCR products were sequenced and compared with 357-bp nucleotide sequences encoding the CEV P4a protein in GenBank. In four cases of disease outbreak (three common carp breeding facilities and one koi garden pond), CEV detected belonged to genogroup I. In one case (koi garden pond), fish were confirmed as infected with CEV from genogroup II. This work complements data on CEV occurrence in European countries and contributes to a better understanding of the pathways leading to transmission of the virus throughout Europe.     

Developing a predictive model for spinal shock in dogs with spinal cord injury

BackgroundReduced pelvic limb reflexes in dogs with spinal cord injury typically suggests a lesion of the L4‐S3 spinal cord segments. However, pelvic limb reflexes might also be reduced in dogs with a T3‐L3 myelopathy and concurrent spinal shock.Hypothesis/ObjectivesWe hypothesized that statistical models could be used to identify clinical variables associated with spinal shock in dogs with spinal cord injuries.AnimalsCohort of 59 dogs with T3‐L3 myelopathies and spinal shock and 13 dogs with L4‐S3 myelopathies.MethodsData used for this study were prospectively entered by partner institutions into the International Canine Spinal Cord Injury observational registry between October 2016 and July 2019. Univariable logistic regression analyses were performed to assess the association between independent variables and the presence of spinal shock. Independent variables were selected for inclusion in a multivariable logistic regression model if they had a significant effect (P ≤ .1) on the odds of spinal shock in univariable logistic regression.ResultsThe final multivariable model included the natural log of weight (kg), the natural log of duration of clinical signs (hours), severity (paresis vs paraplegia), and pelvic limb tone (normal vs decreased/absent). The odds of spinal shock decreased with increasing weight (odds ratio [OR] = 0.28, P = .09; confidence interval [CI] 0.07‐1.2), increasing duration (OR = 0.44, P = .02; CI 0.21‐0.9), decreased pelvic limb tone (OR = 0.04, P = .003; CI 0.01‐0.36), and increased in the presence of paraplegia (OR = 7.87, P = .04; CI 1.1‐56.62).Conclusions and Clinical ImportanceA formula, as developed by the present study and after external validation, could be useful for assisting clinicians in determining the likelihood of spinal shock in various clinical scenarios and aid in diagnostic planning.     

Impact of plant species and atmospheric CO2 concentration on rhizodeposition and soil microbial activity and community composition

Both plant species and CO₂ concentration can potentially affect rhizodeposition and consequently soil microbial activity and community composition. However, the effect differs based on plant developmental stage. We focused on the effect of three plant species (forbs, grasses, and N₂‐fixers) at an early stage of development on root C deposition and fate, soil organic matter (SOM) mineralization and soil microbial community composition at ambient (aCO₂) and elevated (eCO₂) CO₂ levels. Plants were grown from seed, under continuous ¹³C‐labelling atmospheres (400 and 800 µmol mol^?1 CO₂), in grassland soil for three weeks. At the end of the growth period, soil respiration, dissolved organic C (DOC) and phospholipid fatty acid (PLFA) profiles were quantified and isotopically partitioned into root‐ and soil‐derived components. Root‐derived DOC (0.53 ± 0.34 and 0.26 ± 0.29 µg mL soil solution^?1) and soil‐derived CO₂ (6.14 ± 0.55 and 5.04 ± 0.44 µg CO₂‐C h^?1) were on average two times and 22% higher at eCO₂ than at aCO₂, respectively. Plant species differed in exudate production at aCO₂ (0.11 ± 0.11, 0.10 ± 0.18, and 0.58 ± 0.58 µg mL soil solution^?1 for Plantago, Festuca, and Lotus, respectively) but not at eCO₂ (0.20 ± 0.28, 0.66 ± 0.32, and 0.75 ± 0.15 µg mL soil solution^?1 for Plantago, Festuca, and Lotus, respectively). However, no differences among plant species or CO₂ levels were apparent when DOC was expressed per gram of roots. Relative abundance of PLFAs did not differ between the two CO₂ levels. A higher abundance of actinobacteria and G‐positive bacteria occurred in unplanted (8.07 ± 0.48 and 24.36 ± 1.18 mol%) and Festuca‐affected (7.63 ± 0.31 and 23.62 ± 0.69 mol%) soil than in Plantago‐ (7.04 ± 0.36 and 23.41 ± 1.13 mol%) and Lotus‐affected (7.24 ± 0.17 and 23.13 ± 0.52 mol%) soil. In conclusion, the differences in root exudate production and soil respiration are mainly caused by differences in root biomass at an early stage of development. However, plant species evidently produce root exudates of varying quality affecting associated microbial community composition.     

Long-term peroral administration of bitter apricot seeds influences cortical bone microstructure of rabbits

Apricot seeds due to the presence of cyanogenic glycoside amygdalin belong to the popular “alternative cancer cures", although anticancer effect of amygdalin remains controversial. This in vivo study points to the effect of long-term peroral administration of bitter apricot seeds on bone microstructure of rabbits since chronic amygdalin toxicity in relation to bone parameters has not been investigated yet. Rabbits (n = 16) were randomly divided into four experimental groups of 4 animals each. Three experimental groups S1, S2 and S3 received commercial feed for rabbits mixed with crushed bitter apricot seeds at doses 60, 300 and 420 mg/kg bw during five months, respectively. The control (C) group received no apricot seeds. The long-term consumption of apricot seeds had no impact on total body weight, femoral weight and femoral length of rabbits. Also, microcomputed tomography (3D analysis) of cortical and trabecular bone tissues did not reveal any significant impact of amygdalin toxicity on relative bone volume, BMD, cortical bone thickness, bone surface, trabecular number, thickness, and their separation. On the other hand, histological (2D) analysis demonstrated evident changes in cortical bone microstructure consistent with a decreased density of secondary osteons in the middle part of substantia compacta due to a replacement of Haversian bone tissue by plexiform bone tissue, vasoconstriction in the primary osteons' vascular canals, Haversian canals, and decreased sizes of secondary osteons in rabbits from S1, S2 and S3 groups. These negative changes are associated with different vascularization and biomechanical properties of cortical bones.     

Impact of Aluminum on Phosphate Uptake and Acid Phosphatase Activity in Root Tips of Lotus Japonicus

The effect of aluminum (Al) on phosphate homeostasis and induction of acid phosphatase activity and isoenzymes was analyzed in root tips of Lotus japonicus. The rapid uptake of phosphate as well as the decrease of soluble inorganic phosphate in tissues suggested, that the presence of Al in root apoplast causes precipitation of the most of absorbed phosphate. Acid phosphatase (ACP) isoenzymes are affected differentially during low pH and Al stress and both Al induced increases and decreases of ACP isoenzyme activities were observed.     

Resveratrol metabolites do not elicit early pro-apoptotic mechanisms in neuroblastoma cells

Resveratrol, a nontoxic polyphenol, has been shown to inhibit tumor growth in a xenograft mouse model of neuroblasoma. However, resveratrol is rapidly metabolized, mainly to its glucuronidated and sulfated derivatives. This study demonstrates that resveratrol alone, and not the glucuronidated or sulfated metabolites, is taken up into tumor cells, induces a rise in [Ca(2+)](i), and ultimately leads to a decrease in tumor cell viability. A new water-soluble resveratrol formulation was delivered directly at the site of the tumor in a neuroblastoma mouse model. The amount of unmodified resveratrol associated with the tumor increased more than 1000-fold. The increase of unmodified resveratrol associated with the tumor resulted in tumor regression. The number of residual tumor cells that remained viable also decreased as the ratio of the metabolites relative to unmodified resveratrol declined.     

Characterization of adipose-derived equine and canine mesenchymal stem cells after incubation in agarose-hydrogel

Adult stem cells are of particular interest for the therapeutic approach in the field of regenerative medicine. Due to their ease of harvest, adipose-derived mesenchymal stem cells (ASCs) are an attractive stem cell source that has become increasingly popular. Critical aspects of applied cell therapies are the circumstances of transport from the laboratory towards the site of operation and cell delivery into the desired area. With regard to these issues, agarose-hydrogel was analyzed as a cell carrier matrix of equine and canine ASCs in vitro, which can be used for minimally invasive application. Isolated ASCs were expanded and 2.5 × 10⁶ cells were combined with agarose-hydrogel to build a 0.4% hydrogel-cell solution which was stored at two temperatures (room temperature (RT) vs. 37°C). Cell viability was investigated (live-dead assay) at different time points (0, 1, 6 and 24 h) in order to determine i) the effect of different temperatures on the cell survival as well as ii) the maximum possible time span before implantation. CFU-assay and WST-1 assay were performed after 24 h incubation in agarose-hydrogel and the cells were induced into adipogenic and osteogenic differentiation to analyze the effects of the incubation on the cell behaviour. No negative effect of the agarose-hydrogel incubation was determined on the different species’ cell behaviour at either RT or 37°C with any of the assays used. We can recommend agarose-hydrogel as a cell carrier for cell implantation with a storage period of up to 24 h at room temperature or at 37°C prior to implantation.     

Variations on brain microglial gene expression of MMPs, RECK, and TIMPs in inflammatory and non-inflammatory diseases in dogs

Matrix metalloproteinases (MMPs), MMP inhibitors (TIMPs, tissue inhibitors of matrix metalloproteinases), and the membrane-anchored glycoprotein RECK (reversion-inducing cysteine-rich protein with Kazal motifs) contribute to the pathogenesis of many CNS diseases. To assess the potential pathogenetic roles of microglial MMP, TIMP, and RECK generation in extracellular matrix breakdown, opening of the blood brain barrier (BBB) and subsequent recruitment of leukocytes in the CNS, twenty-four dogs suffering from spontaneously occurring different intracranial and extracranial (control group) diseases were examined. Microglia cells were isolated ex vivo by density gradient centrifugation and their expressions of MMP-2, MMP-9, MMP-12, MMP-13, MMP-14, TIMP-1, TIMP-2, and RECK were examined via quantitative real-time polymerase chain reaction (qPCR). Zymography on CNS tissues in selected cases was performed to assess differences at the protein level. Dogs were grouped in different disease categories according to histopathological examinations, in groups with or without inflammatory reactions, and in groups with/without contrast enhancement in advanced diagnostic imaging as a function of BBB breakdown. The results showed a significant up-regulation of MMP-9 in dogs with inflammation in the nervous system compared to dogs with non-inflammatory diseases. An increased expression of MMP-9 might lead to a facilitated invasion of white blood cells. Furthermore, down-regulation of MMP-13 was found in dogs with contrast enhancement. Zymographical data reflected MMP-2 qPCR data. In conclusion, differential expression of MMPs and their inhibitors, but not of RECK, which might crucially influence the pathogenesis of a given disease, could be demonstrated in canine microglia. This reflects a further pathway in the microglial repertoire to respond to various disease conditions in the CNS, a characteristic that might be of particular relevance as a target for specific treatments.     

Effect of long-term feeding of graded levels of deoxynivalenol (DON) on growth performance,nutrient utilization,and organ health in finishing pigs and DON content in biological samples

The prevalence of deoxynivalenol (DON) is a concern for swine producers, and although there has been extensive research into the effects of DON in pigs, focus has been in young pigs and/or in short-term studies. The objective of the study was to determine the effect of long-term exposure to DON-contaminated diets in finisher pigs. A total of 200 pigs (76.6 ± 3.9 kg initial weight) were group housed (five pigs per pen; n = 10 pens/treatment) in a 6-wk trial. Pigs were fed a wheat-barley-soybean meal-based control (CONT) diet with no DON or the basal diet in which clean wheat was replaced by DON-contaminated wheat and wheat screenings to provide DON content of 1, 3, or 5 ppm (DON1, DON3, and DON5, respectively). Individual BW and pen feed intake were recorded weekly to calculate average daily gain (ADG), average daily feed intake (ADFI), and gain to feed ratio (G:F). Blood was collected on days 0, 14, and 43 and analyzed for indicators of liver and kidney health. Nitrogen (N)-balance was conducted immediately following the growth performance period to determine the effect of DON on nutrient utilization. Blood and urine samples collected during N balance were analyzed for DON content. Feeding DON reduced (P < 0.05) ADFI and ADG from days 0 to 28 compared with CONT, after which there was no effect of diet on ADFI and ADG. The G:F was lower (P < 0.05) in DON5 fed pigs compared with all treatments during days 0 to 7; however, no treatment effects on G:F was observed from days 8 to 42. Nitrogen retention was lower (P < 0.05) in DON3 and DON5 compared with DON1-fed pigs. Nitrogen retention efficiency was higher (P < 0.05) in DON1 compared with DON3 and DON5 and protein deposition for DON1 pigs was higher (P < 0.05) than all treatments. There were no treatment effects on indicators of liver and kidney health. As dietary DON intake increased, concentration of DON in blood and urine increased. Overall, although there was an initial decrease in ADG and ADFI in pigs receiving diets containing >1 ppm DON, pig performance recovered after a period of time, whereas nutrient utilization continued to be affected after recovery of performance. Moreover, the lack of DON on G:F indicates that the negative effects of DON on growth performance are largely due to reduced feed intake. Overall, although pigs maybe capable of adapting to intake of DON-contaminated diets, their final body weight will be reduced when fed diets containing >1 ppm DON.