Polyarthritis Caused by Acinetobacter kookii in a Rothschild's Giraffe Calf (Giraffa camelopardalis rothschildi).

We report the first isolation of Acinetobacter kookii from a Rothschild's giraffe calf (Giraffa camelopardalis rothschildi) that had severe polyarthritis. The isolate was resistant to more than one representative of each of four classes of antibiotics (penicillins, macrolides, lincosamides and tetracyclines). As A. kookii has not been previously associated with disease in humans or animals, it may be an emerging opportunistic pathogen posing a threat to immunocompromised patients. Furthermore, as transmission of Acinetobacter spp. with similar patterns of antimicrobial resistance has been previously reported in human and animal populations, special care should be taken when handling infected animals.

Influence of probiotic and yeast culture supplementation on selected biochemical and immunological parameters of growing lambs.

This study was carried out to evaluate the potential effects of 90 days-long dietary supple- mentation of probiotic and yeast culture on immunity condition of lambs. Fifteen Rahmani growing male lambs (about 5 months old and 23.21±2.75 kg body weight) were randomly allo- cated to three equal groups consisting of 5 animals each. The animals in the first group, served as a control (group C), were fed a basal diet without any supplementation. The lambs in the second and third group were fed the basal diet supplemented with probiotic (group Y) or yeast culture (group YC), respectively. The probiotic consisted of live yeast (Saccharomyces cerevisae) alone, while the yeast culture was composed of Saccharomyces cerevisiae and the media on which it was grown. In group Y and YC, each lamb was supplemented daily with 0.5 g and 7.0 g of live yeast and yeast culture, respectively. Blood samples were collected before feeding the supplements and then every 15 days until the day 90th. Total and differential leucocytic counts, total protein, albumin, IgA, IgG and IgM levels were measured in blood. There were insignificant (p>0.05) variations in the levels of total and differential leucocytic counts and total protein among the groups throughout the experiment. However, significant differences (p⟨0.05) were found in globulin, IgA, IgG and IgM in both (Y) and (YC) groups, but the effect of yeast culture seems to be better than that of the probiotic. In conclusions, the obtained results indicate that the tested probiotic and yeast culture improve the immunological status of lambs.

Coagulology, biochemical profile and muscle pathology in calves diagnosed with nutritional muscular dystrophy.

The aim of this study was to explain the correlations between selenium deficiency, hemostatic and biochemical disorders, and the progression of pathological changes in calves diagnosed with nutritional muscular dystrophy (NMD). The study was performed on 20 calves with supplementation of 8 ml selenium and vitamin E preparation and 20 calves with symptoms of NMD. Blood was sampled from calves aged 5, 12 and 19 days. On day 19, samples of the biceps femoris muscle were collected from 6 animals in each group for histopathological analysis. The following blood parameters were determined: PLT, PT, TT, APTT, fibrinogen and D-dimer concentrations, antithrombin III activity, glucose, selenium and vitamin E concentrations, activity of CK, LDH and GSH-Px. Muscle sections were stained with H&E and HBFP. Platelet counts were significantly lower in calves with symptoms of NMD. No significant differences in coagulation parameters were observed between the groups. Sick calves were diagnosed with hyperglycemia and elevation of CK and LDH activity. Selenium and vitamin E concentrations in the blood serum were significantly lower in the experimental group together with significant drop in GSH-Px activity. Changes characteristic of Zenker's necrosis were observed in a muscle of the sick animals. To our best knowledge this is the first study in which the attempt was made to explain the relationship between selenium deficiency and changes in the coagulation system in ruminants.

Bone-forming cells with pronounced spread into the third dimension in polymer scaffolds fabricated by two-photon polymerization.

The main aim of this work was to stimulate bone-forming cells to produce three-dimensional networks of mineralized proteins such as those occurring in bones. This was achieved by a novel approach using a specific type of mesenchymal progenitor cells (i.e., primary fibroblast cells from human hair roots) seeded on to polymer scaffolds. We wrote polymer microstructures with one or more levels of quadratic pores on to a flexible substrate by means of two-photon polymerization using a Ti-sapphire femtosecond laser focused into a liquid acrylate-based resin containing a photoinitiator. Progenitor cells, differentiated into an osteogenic lineage by the use of medium supplemented with biochemical stimuli, can be seeded on to the hydrophilic three-dimensional scaffolds. Due to confinement to the microstructures and/or mechanical interaction with the scaffold, the cells are stimulated to produce high amounts of calcium-binding proteins, such as collagen type I, and show an increased activation of the actin cytoskeleton. The best results were obtained for quadratic pore sizes of 35 µm: the pore volumes become almost filled with both cells in close contact with the walls of the structure and with extracellular matrix material produced by the cells. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 891-899, 2017.

Factors affecting the success rate of treatment of recumbent dairy cows suffering from hypocalcaemia.

We aimed to investigate the ratio of accompanying diseases in cows suffering from clinical hypocalcaemia and their influence on cure rate. In five veterinary practices in different regions of Germany, all recumbent cows around parturition were included in the study for a period of 1 year. After recording the case history a clinical examination was done and a serum sample was taken to measure the concentrations of calcium and phosphorus magnesium, beta-hydroxybutyrate, total bilirubin, cholesterol, urea, and the activities of ASAT, CK, and GLDH. Only cows with hypocalcaemia entered the statistical analysis. Hypocalcaemia was the major cause of recumbency in cows of the second lactation or elder. Muscle damage was the second frequent diagnose in recumbent cows and the major concomitant disease in hypocalcaemic cows. The overall cure rate was between 89.4% and 94.8%. Calcium and phosphorus concentrations did not have an influence on cure rate. Non-cured cows had higher serum activities of CK (p<0.043) and ASAT (p<0.006). Nevertheless, the activities of CK and ASAT were no good predictors of treatment failure because of their low specificity and the high cure rate of the cows in the five practices.

Intrinsic thermal resistance of the canine brain.

Hyperthermia above a critical threshold results in multisystemic changes that include neurological manifestations of heat stroke. It is unknown if the latter represents an intrinsic thermal sensitivity of the CNS or whether injury is secondary to physiological responses of non-CNS origin. To address this issue, the present work examined functional, structural, and biochemical changes in the CNS of dogs subjected to a thermal dosage immediately below that which induces disseminated intravascular coagulation with secondary multiple organ injury. The experimental approach is previously reported, inducing a 42.5 degrees C, 90 min, whole body hyperthermia while preventing other physiological responses to treatment, including respiratory alkalosis and significant reductions in mean arterial pressure. Functional analyses included neurologic examinations and brainstem auditory evoked potential recordings in the post-treatment interval in both hyperthermic and euthermic control populations. Biochemical and structural analyses examined the expression of 70-kDa heat shock proteins, cytokines, markers of astroglial and microglial injury/activation, evidence of vascular endothelial damage, and evidence of neuronal and axonal injury in brain between 0.5 h and 8 days from the end of the treatment. The only significant change associated with treatment was induction of the major inducible 70-kDa heat shock protein, this being most prominent in the cerebellum with maximal expression at 6 h and a return to baseline by 8 days.Collectively, from these results we suggest that the canine brain is intrinsically resistant to sublethal hyperthermia such that when CNS lesions occur, they do so in the presence of other physiological derangements.

Perimodiolar electrodes in cochlear implant surgery.

Perimodiolar-positioned cochlear implant electrodes have been developed in order to bring the electrode contacts as close as possible to the spiral ganglion cells, which are the target of electrostimulation. This results in lower electrical thresholds, higher dynamic ranges and less channel interaction when compared with normal implant electrodes which are usually located peripherally within the scala tympani. In this study we evaluated 4 different types of perimodiolar electrode: the Clarion Preformed electrode, the Clarion Preformed electrode with positioner, the Nucleus Contour electrode and the Med-El Perimodiolar Combi 40 electrode. These devices require different approaches to achieve a perimodiolar electrode position. The electrodes were inserted in fresh human temporal bones. After processing these bones with the electrodes in situ by employing a sawing, grinding and polishing technique, the inner ear structures as well as the electrode positions could be evaluated in detail. All electrode types studied had a more or less perimodiolar position; however, each type produced a certain amount of trauma to cochlear structures which is discussed in relation to mechanical properties. Further human temporal bone studies with improved perimodiolar cochlear implant electrodes are necessary in order to find an optimized type of electrode.

Multiple isoforms of porcine aromatase are encoded by three distinct genes.

Cytochrome P450 aromatase, a product of the CYP 19 gene and the terminal enzyme in the estrogen biosynthetic pathway, is synthesized by the ovary, endometrium, placenta, and peri-implantation embryos in the pig and other mammals, albeit to varying levels, implying its functional role(s) in pregnancy events. The aromatase produced by the pig tissues exists as three distinct isoforms (type I - ovary, type II - placenta, and type III - embryo), with presumed differences in substrate specificities, expression levels, activity, and mode of regulation. In order to delineate the molecular mechanisms whereby estrogen synthesis is regulated in these diverse tissues, the present study examined if these aromatase isoforms represent products of multiple genes or of a single gene via complex splicing mechanisms. Porcine genomic DNA from a single animal was used as a template in the polymerase chain reaction (PCR) to amplify isoform-specific sequences corresponding to exons 4 and 7, respectively. Nucleotide sequence analysis of the generated fragments revealed the presence of only clones corresponding to the three known aromatase types. Screening a porcine Bacterial Artificial Chromosome (BAC) library for aromatase gene by PCR yielded a single clone approximately 80 kb in length. Southern blot analysis, using probes specific for exons 1A-1B, 2-3, 4-9, and 10 sequences indicated that the BAC genomic clone contains the entirety of the coding exons as well as the proximal promoter region. Sequence analysis of the fragment generated with exon 4 primers determined that this BAC clone contains only the type II gene. The presence and relative orientation of the untranslated 5'- exons 1A and 1B, previously demonstrated for the type III isoform were evaluated in the BAC clone and genomic DNA by PCR. The 265 bp fragment generated from both PCR reactions was confirmed by sequence analysis to contain exons 1A and 1B that are located contiguous to each other and separated by only three bp. A diagnostic procedure for typing aromatase isoforms was developed, based on the presence of specific restriction sites within isoform-specific exons. The use of this protocol confirmed the existence of only three aromatase isoforms in the porcine genome and indicated changes in aromatase types expressed by the uterine endometrium as a function of pregnancy stage. The presence of distinct genes encoding each of the aromatase isoform predicts important differences in the mechanisms underlying the molecular evolution and regulation of porcine aromatase, unique from those of other mammals, and suggests a critical role for P450 aromatase steroidal products in uterine functions related to pregnancy events.

Effects of humoral hypercalcemia of malignancy on the parathyroid gland in nude mice.

The ultrastructure of parathyroid chief cells was examined from four groups of nude mice (NIH:Swiss) with different serum calcium concentrations. The groups consisted of eight male mice with hypercalcemia induced by transplantable canine adenocarcinoma (CAC-8), eight female mice with hypercalcemia induced by infusion of parathyroid hormone-related protein, ten male control mice, and six male mice fed a low calcium (0.01%) diet. Hypercalcemia induced by malignancy or parathyroid hormone-related protein infusion was associated with low serum phosphorus concentration, a decrease in the number of secretory and prosecretory granules in the parathyroid chief cells, and an increase in the cytoplasmic area of chief cells. Prominent myelinlike membranous whorls were present in the cytoplasm of chief cells of tumor-bearing and parathyroid hormone-related protein-infused hypercalcemic mice. Mice fed a low calcium diet had decreases in the number of secretory granules and cell area but increases in the number of prosecretory granules compared with control mice. The number of mitochondria and the nuclear area of chief cells were similar in all four groups. The prominent membranous whorls and increased cytoplasmic area of chief cells from these hypercalcemic mice mark these cells as distinctly different from the parathyroid chief cells of other species with hypercalcemia.

Early phosphorus 31 nuclear magnetic resonance bioenergetic changes potentially predict rejection in heterotopic cardiac allografts.

The development of a noninvasive screening test for the detection of cardiac allograft rejection would improve the potential for management of heart transplant recipients. To assess the possibility that changes in myocardial high-energy phosphate metabolism precede frank rejection, 17 beagles received cervical cardiac allografts. Recipients underwent serial phosphorus 31 nuclear magnetic resonance spectroscopy, endocardial biopsy (blindly graded, 0 to 8), and left ventricular pressure measurements starting on the day of surgery. The first (less than 24 hours) spectrum was considered the baseline for all additional studies. The phosphocreatine to inorganic phosphate ratio (PCr/Pi), an index of myocardial bioenergetic supply/demand balance, was determined and expressed as a percentage of baseline of initial and all subsequent spectra. To evaluate the predictive utility of the PCr/Pi ratio, a 50% decrease from baseline was designated as a positive test and was correlated with biopsy-proved rejection (score greater than 3). When PCr/Pi values were compared with the subsequent day's biopsy score, we observed a 91% sensitivity, 90% specificity, and a predictive value of 92%. We conclude that the PCr/Pi ratio is sensitive in predicting heterotopic allograft rejection in its earliest stages. Thus phosphorus 31 nuclear magnetic resonance holds promise for clinical use in the noninvasive diagnosis and monitoring of cardiac rejection.

31P nuclear magnetic resonance determination of changes in energy state in lung preservation.

The significance of dynamic changes in energy state during lung harvesting and preservation has not been extensively studied. Phosphorus 31 nuclear magnetic resonance spectra at 81 MHz were obtained from degassed rabbit lungs. Changes in the adenosine 5'-triphosphate-to-inorganic phosphate peak-intensity ratios were used to measure changes in energy state. Two groups of rabbit preparations were studied to evaluate the effect of hypothermia during the initial 120 minutes of harvesting (n = 8 at 36 degrees C and n = 5 at 4 degrees C). The significance of these changes was assessed in a second experiment in which lungs were reperfused in vitro at selected intervals of hypothermia (5, 12, and 24 hours) and assessed for injury. Hypothermic preservation sustained a significantly higher energy state. The depletion of energy state was correlated with injury, particularly as measured by lung edema (r2 = -0.715). Short periods of warm ischemia (30 minutes) result in a significant depletion of energy state, which may be a component of pulmonary injury during harvesting and preservation.

Antioxidant effects in the development of Ehrlich ascites carcinoma.

The effect of Ehrlich ascites tumor growth on selenium-turnover rates and selenium-75 distribution in liver, kidney, and immunological tissues (spleen, thymus, and lymph nodes) was investigated in Swiss Webster mice that had been prelabeled with selenium-75. Ehrlich ascites tumor caused a decrease in the selenium-75 content of liver, kidney, and thymus; it also decreased the rate of the total-body selenium-turnover. In liver, depletion of selenium-75 was almost as great as that produced by a selenium and vitamin E-deficient diet. When mice had been fed an antioxidant-deficient diet, considerable quantities of selenium-75 were accumulated by the tumor; the specific activity of the tumor increased 9-fold over that in antioxidant-supplemented mice. The same diet produced a premature, and in some cases drastic, contraction in tumor volume. The possible significance of tumor-induced antioxidant deficiencies to the etiology of certain paraneoplastic syndromes is discussed.